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植物生物化学与分子生物学(中文版)(Plant Biochemistry and Molecular Biology

中文名: 植物生物化学与分子生物学(中文版)原名: Plant Biochemistry and Molecular Biology资源格式: PDF发行时间: 2004年地区: 大陆语言: 简体中文简介:作者：(美)B.B.布坎南(BobB.Buchanan)出版社：科学出版社出版日期：2004年2月版次：ISBN：703012013 页数：1090开本：大16开包装：价格：￥260.0本书简介本书英文版由国际杰出植物生物学家编写，美国植物生物学家学会出版，是植物生物学领域的重要著作。

在整合前沿知识的基础上，本书围绕细胞区室结构、细胞的繁衍、能量流、代谢与发育的整合、植物的环境与农业5个主题精心组织内容，反映了各个领域的研究历史和最新进展。

本书编排有序，图文并茂，适用于植物生物学以及分子生物学、生物技术、生物化学、细胞生物学、生理学、生态学等相关领域的研究和教学参考。

制药学、农业经济等领域的研究人员也可从中得到有价值的信息。

目录:第1篇区室结构1 膜结构和被膜细胞器导言1.1 细胞膜的共性和遗传性1.2 膜的流动镶嵌模型1.3 质膜1.4 内质网1.5 高尔基体1.6 胞吐和内吞1.7 液泡1.8 细胞核1.9 过氧化物酶体1.10 质体1.11 线粒体小结相关文献2 细胞壁导言2.1 糖：组成细胞壁的基本单位2.2 组成细胞壁的大分子2.3 细胞壁构架2.4 细胞壁的生物合成和装配2.5 生长与细胞壁2.6 细胞分化2.7 可用作食物、饲料和纤维的细胞壁小结相关文献3 膜转运导言3.1 膜运输概述3.2 植物膜上运输的组织构成3.3 泵3.4 载体蛋白3.5 离子通道的一般特性3.6 运转中的离子通道3.7 通过水通道蛋白运输水小结相关文献4 蛋白质分选和囊泡运输导言4.1 蛋白质分选的机制4.2 将蛋白质定位到质体中4.3 转运进入线粒体和过氧化物酶体4.4 细胞核的内向和外向转运4.5 内质网在蛋白质分选和组装中的作用4.6 液泡定位和分泌4.7 高尔基体中的蛋白质修饰4.8 内吞作用小结相关文献5 细胞骨架导言5.1 细胞骨架概述5.2 中间纤维5.3 肌动蛋白与微管蛋白家族5.4 肌动蛋白与微管蛋白的聚合5.5 肌动蛋白与微管蛋白的特性5.6 细胞骨架结合蛋白5.7 肌动蛋白纤维在胞内定向运动中的作用5.8 皮层微管与细胞扩展5.9 观察细胞骨架的动力学5.10 细胞骨架与信号转导5.11 细胞骨架与有丝分裂5.12 细胞骨架与胞质分裂小结相关文献第2篇细胞的繁衍6 核酸导言6.1 核酸的组成与核苷酸的合成6.2 细胞核DNA的复制6.3 DNA修复6.4 DNA重组6.5 细胞器DNA6.6 DNA转录6.7 RNA的特征和功能6.8 RNA加工小结相关文献7 基因组的组织结构与表达导言7.1 基因与染色体7.2 核基因组的组织结构7.3 转座因子7.4 基因表达7.5 染色质在染色体组织和基因表达中的作用7.6 基因调控的后生遗传机制小结相关文献8 氨基酸导言8.1 植物体内氨基酸的生物合成：研究现状与前景8.2 无机氮同化进N-转运氨基酸8.3 芳香族氨基酸的合成8.4 天冬氨酸衍生氨基酸的生物合成8.5 支链氨基酸8.6 脯氨酸代谢：耐胁迫代谢工程的靶标小结相关文献9 蛋白质的合成、装配和降解导言9.1 从RNA到蛋白质9.2 真核生物细胞质蛋白质生物合成的调控9.3 叶绿体中蛋白质的合成9.4 蛋白质的翻译后修饰9.5 蛋白质降解相关文献10 脂类导言10.1 脂类的结构与功能10.2 脂肪酸的生物合成10.3 乙酰辅酶A羧化酶10.4 脂肪酸合酶10.5 C16 和C18 脂肪酸的去饱和及其延长10.6 特殊脂肪酸的合成10.7 膜脂的合成10.8 膜脂的功能10.9 结构脂类的合成与功能10.10 贮藏性脂类的合成与分解代谢10.11 脂类的基因工程小结相关文献11 细胞分裂的调控导言11.1 动植物细胞及其细胞周期11.2 细胞周期研究的历史回顾11.3 DNA复制11.4 有丝分裂11.5 细胞周期的调控机制11.6 细胞周期的调控逻辑11.7 多细胞生物的细胞周期调控11.8 植物生长发育中的细胞周期调控小结相关文献第3篇能量流12 光合作用导言12.1 光合作用总论12.2 光吸收与能量转换12.3 反应中心复合体12.4 光系统12.5 类囊体膜的组成12.6 叶绿体膜的电子转移途径12.7 叶绿体中的A TP合成12.8 C3植物中的碳反应12.9 CO2固定机制的差异小结相关文献13 糖代谢13.1 磷酸己糖库13.2 利用磷酸己糖的生物合成途径：蔗糖和淀粉的合成13.3 产生磷酸己糖的分解代谢途径：蔗糖和淀粉的降解13.4 磷酸丙糖/磷酸戊糖代谢产物库13.5 磷酸己糖和磷酸戊糖/磷酸丙糖代谢产物库之间的相互作用13.6 淀粉与蔗糖合成：细胞对代谢总调控的范例13.7 糖类对基因表达的调控13.8 糖酵解中的贮能反应13.9 为生物合成反应提供能量和还原力小结相关文献14 呼吸与光呼吸导言14.1 呼吸概论14.2 柠檬酸(三羧酸)循环14.3 植物线粒体的电子传递14.4 植物线粒体的ATP合成14.5 线粒体呼吸作用的调节14.6 线粒体与细胞其他区域的相互关系14.7 光呼吸的生化基础14.8 光呼吸途径14.9 植物中光呼吸的规律小结相关文献第4篇代谢与发育的整合15 长距离运输导言15.1 植物体内物质的扩散与径流15.2 通道大小在确定质外体和共质体运输特征中有重要作用15.3 木质部和韧皮部物质运输的比较15.4 木质部中水分的蒸腾运动15.5 胞间连丝介导的共质体运输15.6 韧皮部运输15.7 植物内源大分子的细胞间运输小结相关文献16 氮和硫导言16.1 生物圈和植物中氮素概况16.2 固氮概论16.3 氮固定中的酶学16.4 共生固氮16.5 氨的吸收和运输16.6 硝酸盐的吸收和还原概述16.7 硝酸盐的还原16.8 亚硝酸盐的还原16.9 硝酸盐同化和碳代谢间的相互作用16.10 硫酸盐同化概述16.11 硫的化学性质及功能16.12 硫的吸收及运输16.13 还原硫的同化途径16.14 谷胱甘肽及其衍生物的合成及功能小结相关文献17 植物激素与诱激物分子的生物合成导言17.1 赤霉素17.2 脱落酸17.3 细胞分裂素17.4 吲哚-3-乙酸17.5 乙烯17.6 油菜素类固醇17.7 多胺17.8 茉莉酮酸17.9 水杨酸17.10 展望小结相关文献18 信号感受和转导导言18.1 信号转导概述18.2 受体18.3 植物受体的特殊例子18.4 G蛋白和磷脂信号系统18.5 环状核苷酸18.6 钙18.7 蛋白激酶：信号转导中的基本组分18.8 植物生长调节因子参与特殊的信号转导途径18.9 植物细胞信号转导研究的展望小结相关文献19 生殖发育导言19.1 开花诱导19.2 花的发育19.3 花发育的遗传和分子分析19.4 配子的形成19.5 影响配子体发育的突变19.6 花粉的萌发19.7 自交不亲和19.8 受精作用19.9 种子形成19.10 种子发育过程中贮藏物质的积累19.11 胚胎的成熟和脱水19.12 萌发小结相关文献20 衰老与程序性细胞死亡导言20.1 动物及植物中观察到的细胞死亡的类型20.2 植物生活周期中的PCD20.3 衰老概述20.4 衰老过程中的色素代谢20.5 衰老过程中的蛋白质代谢20.6 衰老对光合作用的影响20.7 衰老对氧化代谢的影响20.8 衰老过程中的核酸降解20.9 衰老细胞中代谢活性的调节20.10 内源植物生长调节因子与衰老20.11 环境对衰老的影响20.12 植物发育性PCD的例子：管状分子的形成和禾本科植物内胚乳的转移20.13 PCD作为植物胁迫应答的例子：通气组织的形成和超敏反应20.14 PCD研究的未来方向以及面临的更多问题小结相关文献第5篇植物的环境与农业21 植物对病原体的反应导言21.1 植物病原体的致病机理21.2 植物防御系统21.3 植物－病原体相互作用的遗传基础21.4 R基因与R基因介导的植物抗病性21.5 植物防御反应的生化原理21.6 系统性植物防御反应21.7 利用基因工程控制植物病原体小结相关文献22 植物对非生物胁迫的反应导言22.1 植物对非生物胁迫的反应22.2 与缺水相关的胁迫22.3 渗透调节及其在耐旱耐盐中的作用22.4 缺水和盐分对跨膜转运的影响22.5 水分胁迫诱导的其他基因22.6 冰冻胁迫22.7 水涝和缺氧22.8 氧化胁迫22.9 热胁迫小结相关文献23 矿质营养吸收、转运及利用的分子生理学导言23.1 必需矿质元素概论23.2 植物K+转运机制与调节23.3 磷的营养与转运23.4 微量营养吸收的分子生理学23.5 植物对矿质毒性的反应小结相关文献24 天然产物(次生代谢物)导言24.1 萜类化合物24.2 IPP的生物合成24.3 异戊烯转移酶与萜类合酶参与的反应24.4 萜类化合物骨架的修饰24.5 转基因萜类产物24.6 生物碱24.7 生物碱的生物合成24.8 生物技术在生物碱生物合成研究中的应用24.9 苯丙烷类化合物和苯丙烷类－乙酸酯途径的代谢产物24.10 苯丙烷类化合物和苯丙烷类－乙酸酯的生物合成24.11 木脂体、木质素的生物合成和栓化作用24.12 黄酮类化合物24.13 香豆素、芪、苯乙烯吡喃酮和芳基吡喃酮类化合物24.14 苯丙烷类产物的代谢工程：改善纤维、色素、药物和调味剂的可能途径小结相关文献索引。

分子生物学中的基因转录和翻译

分子生物学中的基因转录和翻译基因是生命的基本单位，是人类、动物和植物的遗传信息载体。

基因可以转录为RNA，并且RNA可以被翻译为蛋白质。

基因转录和翻译是维持细胞和生物体正常生理功能的重要过程。

基因转录基因转录是指DNA水平上的信息传递，即将DNA编码的信息转换为RNA信息，并用来推断蛋白质的氨基酸序列。

基因转录是由RNA聚合酶(RNA polymerase)复制DNA时合成RNA分子的过程，RNA聚合酶会在DNA串内扫描，寻找一段特定的DNA序列，其通常以一个起始站点开始，称为启动子。

在这个地方，RNA聚合酶结合并开始克隆RNA。

这个启动序列通常是由两个特定的功能元件组成。

第一部分是TATA盒(TATA box)，它告诉RNA聚合酶在哪里开始转录。

第二部分是增强子(enhancer)序列，它可以增加基因的表达并协调DNA复制的过程。

完成转录之后，pre-mRNA序列会被剪切并拼接，形成成熟的mRNA。

mRNA可以被转运到细胞质中并参与翻译过程。

转录的主要产物是mRNA，但是转录也可以产生其他类型RNA。

转录的调控是生物体中基因表达的关键控制因素。

细胞可以通过控制RNA聚合酶与DNA的互作、核糖体合成和RNA降解等因素来控制基因转录的发生。

此外，转录的调控还受到一些核酸因子和转录激活因子的影响。

许多疾病，如肿瘤和自身免疫疾病，都与转录调控紊乱有关。

基因翻译基因翻译是指RNA水平上的信息传递，即通过将RNA信息翻译为氨基酸序列，生成蛋白质。

蛋白质质量和结构的确定取决于氨基酸的顺序。

20种不同的氨基酸可以以不同的序列组合来进一步分别形成不同的蛋白质。

蛋白质的信息来源于mRNA，mRNA中通过第三个核苷酸测序，信息被读取为三个核苷酸组成的非重叠密码子的序列。

在翻译过程中，一个RNA分子会通过核糖体与一个氨基酸专一地配对，然后一个又一个的氨基酸加入到正在被构建的多肽链中。

翻译是一个复杂的过程，它涉及到许多因素，如翻译起始和停止位点的识别、翻译调节和后翻译修饰等。

分子生物学词汇(中英文对照表 )

分子生物学中的转录和翻译过程

分子生物学中的转录和翻译过程转录和翻译是分子生物学中的两个重要过程。

转录是指从DNA模板合成RNA分子的过程，其中RNA作为信息的中介传递到细胞内的核外，然后供翻译使用。

翻译是指将RNA翻译成蛋白质序列的过程，是生命体系中产生多种功能蛋白质的基础。

本文将分别介绍这两个过程的机制和重要性。

一、转录过程转录是一种基因表达过程，它涉及到模板DNA的开放和RNA合成。

本质上，转录是一种DNA依赖性RNA合成过程，能够启动生物体内大多数核苷酸序列的表达。

相比DNA，RNA分子更易于合成和分解，并且具有许多不同类型：传递RNA（tRNA）、转运RNA（rRNA）和信使RNA（mRNA）等。

转录过程的主要步骤如下：1. 启动子序列的结合：RNA聚合酶必须与某种DNA序列结合才能启动合成RNA的过程。

启动子序列通常位于基因的起始位置，用于指示RNA酶具体在哪一片段开始转录。

2. 开链：RNA酶从DNA双链中打开某一区段，从而产生一个开放的DNA单链。

该单链被稳定地保护，以避免在转录期间被其他元件损坏。

3. 合成RNA：RNA聚合酶沿着单链DNA向前移动，并利用进入口处的核苷酸再合成一个反义核苷酸链的RNA分子。

RNA聚合酶仅将核苷酸添加到5'末端，仅被用作RNA合成起始部分的碱基标志在3'末端停止合成。

整个过程持续到RNA合成末端的终止序列，然后RNA成品释放，并RNA聚合酶从DNA模板中离开。

二、翻译过程翻译是将RNA序列转化为蛋白质的序列的过程，可以分为三个主要步骤：启动、延长和终止。

启动从AUG（起始）密码子开始，在三联码（一种由三个核苷酸组成的密码子，每个三联码都代表一条氨基酸）的作用下继续进行。

翻译过程必须稍微转换一下信息：DNA中的碱基序列被翻译成RNA中的天然核苷酸单元，然后转变为氨基酸的多肽链中的化学信号。

然而，在许多细胞中，许多会影响翻译机制的复杂调节机制也存在。

三、结论转录翻译是基因表达的重要过程，可实现生命中原始信息的继承、分化和增加。

分子生物学chapter1绪论

大量生产某些正常细胞代谢中产量很低的多肽，如激素、抗生素、酶类及抗体等，提高产量，降低成本。
定向改造某些生物的基因组结构、使它们所具有的特殊经济价值或功能得以成百上千倍地提高。
进行基础研究。
2、基因表达调控研究
蛋白质分子控制了细胞的一切代谢活动，而决定蛋白质结构和合成时序的信息都由核酸（主要是脱氧核糖核酸）分子编码，所以，基因表达实质上就是遗传信息的转录和翻译过程。
基本定理：
1.构成生物体有机大分子的单体在不同生物中都是相同的；
2.生物体内一切有机大分子的建成都遵循着各自特定的规则；
3.某一特定生物体所拥有的核酸及蛋白质分子决定了它的属性。
1、DNA重组技术
是20世纪70年代初兴起的技术科学，目的是将不同DNA片段（基因或基因的一部分）按照人们的设计定向连接起来，在特定的受体细胞中与载体同时复制并得到表达，产生影响受体细胞的新的遗传性状。
DNA重组技术是核酸化学、蛋白质化学、酶工程及微生物学、遗传学、细胞学长期深入研究的结晶，而限制性内切酶DNA连接酶及其他工具酶的发现与应用则是这一技术得以建立的关键。
通过DNA连接酶把不同的DNA片段连接成一个整体。a. DNA的粘性末端; b. DNA的平末端; c. 化学合成的具有EcoRI粘性末端的 DNA片段。
1．拥有特定的空间结构（三维结构）；
2．在它发挥生物学功能的过程中必定存在着结构和构象的变化。
4、基因组、功能基因组与生物信息学研究
2001年2月，Nature 和Science同时发表了人类基因组全序列。
已有数十种生物基因组被基本破译。
测定基因组序列只是了解基因的第一步，在基因组计划的基础上提出了蛋白质组计划（又称后基因组计划或功能基因组计划），旨在快速、高效、大规模鉴定基因的产物和功能。

专业英语翻译：分子生物学词汇

专业英语翻译：分子生物学词汇convalescent phase serum 恢复期血清convallamarin 铃兰苦苷convallaria cardiac glycoside 铃兰（类）强心苷convallarin 铃兰苷convective mass transfer 对流传质convective transfer 对流传递conventional mouse 常规小鼠convergence 会聚[用于神经系统]；趋同convergent evolution 趋同进化convergent synthesis 汇集合成conversion （基因）转变cooked meat medium 庖肉培养基cooling 冷却cooling air 冷却空气cooling bath 冷却水浴cooling coil 冷却旋管cooling jacket 冷却套管，冷却夹套cooling system 冷却系统cooling tower 冷却塔cooling tube 冷却管cooling water 冷却水cooling water circulation 冷却水循环coomassie blue [商]考马斯蓝coomassie brilliant blue [商]考马斯亮蓝cooperation 协同（作用）cooperative 协同的cooperative effect 协同效应，合作效应cooperative feedback inhibition 协同反馈抑制cooperative site 协同部位，协同位点cooperativity 协同性coordinate 坐标coordinate axis 坐标轴coordinate bond 配位键coordinate regulation 协同调节coordination 配位（作用）；协同（作用），协调（作用）coordination agent 配位剂coordination anion 配（位）阴离子coordination bond 配位键coordination catalysis 配位催化coordination cation 配（位）阳离子coordination compound 配位化合物，配合物coordination ion 配离子coordination isomerism 配位异构coordination number 配位数coordination site 配位点coordination sphere 配位层copolycondensation 共缩聚copolymer （二元）共聚物copolymerization 共聚合（反应）copper grid 铜载网copper protein 铜蛋白[共包括i,ii、iii型，i型即铜蓝蛋白] coprecipitation 共沉淀coprinin 鬼伞菌素coproporphyrin 粪卟啉coproprophyrinogen 粪卟啉原coprosterol 粪固醇，粪甾醇copurification 共纯化copy error 复制错误copy number 拷贝数cord factor 索状因子[如见于白喉杆菌]cordycepin 3'-脱氧腺苷；蛹虫草菌素。

各专业课程英文翻译

各专业课程英⽂翻译各专业课程英⽂翻译（精⼼整理）⽣物及医学专业课程汉英对照表细胞⽣物学和分⼦⽣物学 Celluar and Molecular Biology 精神病护理学 Psychiatric Nursing⼒学专业⾼等代数与⼏何 Advanced Algebra and Geometry 数学物理⽅法 Methods in Mathematical Physics理论⼒学 Theoretical Mechanics 弹性⼒学 Elasticity⼒学实验 Experiments in Solid Mechanics ⼒学概论Introduction to Mechanics 计算流体⼒学 Computational Fluid Mechanics 粘性流体⼒学Viscous Fluid Flow弹性⼒学变分原理 Variational Principles inElasticity有限元法 Finite Element Method进化⽣物学 Evolutionary Biology ⼝腔外科学 Oral Surgery海洋⽣物学 Marine Biology ⼝腔 / ⽛科科学 Oral/Dental Sciences 微⽣物学 Microbiology⾻科医学 Osteopathic Medicine 分⼦⽣物学 Molecular Biology ⽿科学 Otology医学微⽣物学 Medical Microbiology 理疗学 Physical Therapy ⼝腔⽣物学 Oral Biology ⾜病医学Podiatric Medicine寄⽣物学 Parasutology眼科学 Ophthalmology 植物⽣物学 Plant Physiology 预防医学Preventive Medicine ⼼理⽣物学 Psychobiology 放射学 Radiology放射⽣物学 Radiation Biology 康复咨询学 Rehabilitation Counseling 理论⽣物学 Theoretical Biology 康复护理学Rehabilitation Nursing 野⽣⽣物学 Wildlife Biology 外科护理学 Surgical Nursing 环境⽣物学 Environmental Biology 治疗学Therapeutics 运动⽣物学 Exercise Physiology 畸形学 Teratology有机体⽣物学 Organismal Biology 兽医学 Veterinary Sciences ⽣物统计学 Biometrics ⽛科卫⽣学 Dental Sciences ⽣物物理学 Biophysics ⽛科科学 Dentistry ⽣物⼼理学 Biopsychology ⽪肤学 Dermatology ⽣物统计学 Biostatistics 内分泌学Endocrinology ⽣物⼯艺学 Biotechnology 遗传学 Genetics ⽣物化学 Biological Chemistry解剖学 Anatomy⽣物⼯程学 Biological Engineering ⿇醉学 Anesthesia ⽣物数学 Biomathematics临床科学 Clinical Science应⽤⽣物学 Applied Biology 细胞⽣物学 Cell Biology ⽣物学 Biology⽣物医学科学 Biomedical Science临床⼼理学 Clinical Psychology 医学技术 Medical Technology 医学 Medicine护理⿇醉学 Nurse Anesthesia 数学分析 Mathematical Analysis 常微分⽅程 Ordinary Differential Equation 计算⽅法Numerical Methods 材料⼒学 Mechanics of Materials 流体⼒学Fluid Mechanics 机械制图 Machining Drawing ⽓体⼒学 Gas Dynamics 弹性板理论 Theory of Elastic Plates塑性⼒学Introduction of Plasticity经典⼒学中的数学⽅法 Mathematical Methods of ClassicalMechanics机器⼈动⼒学 Dynamics of Robots ⾃动控制原理 Principles of Automatic Control优化计算与最化控制 Optimization and OptimalControl计算机图形学 Computer Graphics 概率与统计 Probability and Statistics 专业英语 English for Mechanics 振动理论 Theory of Vibration 程序设计⽅法 (C 和 FORTRAN) Programming in C & FORTRAN ⽔动⼒学 Hydrodynamics 计算机图象处理 Image Processing 光测⼒学 Photo Mechanics断裂⼒学 Fracture Mechanics⾼等动⼒学 Advanced Dynamics 摄动⽅法 Perturbation Methods 机械设计与 Auto CADMachinery Designing and AutoCAD 信息显⽰ (可视化 ) Visualization 微机原理 Principles of Personal Computer 复变函数Complex Function企业管理专业微观经济学 Microeconomics管理信息系统 Systems of Management Information 财务管理 Financial Management 战略管理 Strategic Management 国际商务谈判 Negotiation on BusinessAffairs跨国公司专题研究 Special Researchof multinational corporation国际贸易 InternationalTrade 国际营销研究 International Marketing Research 公司组织与管理 Organization and Managementof Corporate信息管理概论 Introduction to InformationManagement信息经济学 Information Economics 企业信息化⼯程管理学原理 Principles of Management 信息政策与法规 Information Policy and Law 管理信息系统 Management Information System 线性代数 Linear Algebra决策分析 Policy Making 离散数学 Discrete Mathematics概率统计 Statistics and ProbabilityTheory ⽣产与运作管理 Production Management 电⼦商务Electronic Commerce 信息系统安全与保密 Information System Security政府信息化⼯程 Government Informationalization ⼴告实务 Practice of Advertisement 操作系统 Operating System 信息科学基础Foundations of InformationScience 经济信息管理 Economic Information Management 专业英语 Specialty English 微机基础Principles of Microcomputers ⽂献计量学 Bibliometrics电⼦出版技术 Electronic Publishing ⼴告概论 Introduction to Advertisement管理学 Principles of Management 宏观经济学 Macroeconomics 产业经济学 Industrial Economics 项⽬评估 Projects Appraisal 管理沟通Management Negotiation战略管理 Strategic Management ⽣产管理研究 Operation Management 企业伦理 Enterprise Ethics 运筹学 Operational Research 信息管理专业⾼等数学 Higher Mathematics数据库系统 Database 组织⾏为学 Organizational Behavior⼈⼒资源管理 Human Resource Management信息存储与检索信息服务与Information Retrieval andStorage information Service and UserStudy社会实践 Practical Work信息分析与决策 Information Analysis andPolicy Making Enterprise Informationalization 信息组织 Information Organization 计算机⽹络 Computer Networks 多媒体技术 Multimedia信息环境论 Information Environments 传播学原理 Principles of CommunicationTheory 知识产权法学 Law of Intelligence Property 组织⾏为学 Studies of Organization 货币银⾏学专业货币银⾏学 Money and Banking 宏观经济学 Macroeconomics 策略管理 Strategic Management 银⾏会计 Bank Accounting 运筹学 Operational Research 财务管理Financial Management 租赁与信托 Hiring and Affiancing 商业银⾏实务 Practice of Business Bank项⽬评估 Projects Appraisal ⾦融市场学⼈⼒资源管理 Human ResourceManagement 财务报告分析 A nalysis of Financial Statement财务案例分析 Case Analysis of FinancialManagement物理专业热学 Thermodynamics ⼒学 Mechanics光学 Optics电磁学 Electromagnetism计算概论 Computing Generality 普通物理实验 General Physics Laboratory固体磁性及应⽤基础 Magnetism of the Solid Stateand its Application衍射物理(固体结构分析)Diffraction Physics (Structureof Solid Analysis)科研实⽤软件 Utility Software for ScientificResearch 计算机模拟⽅法 Computer SimulationMethods 激光原理、技术与应⽤ The Principle, Techniqueand Application of Laser材料物理 Materials Physics 近代光学和光电⼦学 Modern Optics and Optoelectronics现代固体物理 Modern Solid State Physics粒⼦物理 Particle Physics 物理宇宙学基础 Elements of Cosmology Physics 固体物理 Solid State Physics 原⼦物理 Atomic Physics量⼦⼒学 Quantum Mechanics 理论⼒学 Theoretical Mechanics电动⼒学 Electrodynamics普通物理综合实验 Synthetical Experiments ofGeneral Physics市场营销学专业营销管理 Marketing Management 公共关系 Public Relationship 国际贸易 International Trade 消费者⾏为 Consumer Behavior 管理信息系统 Systems of Management Information 营销调研Marketing Research 推销学 Sales Strategies 国际⾦融 International Finance 营销预测与规划 Marketing Forecasting andPlanning 销售渠道管理 ales Channels Management 管理学 Principles of Management 国际市场营销 International Marketing 商业谈判 Business Negotiation ⼴告管理 Advertising Management 营销案例分析 Case Studies of Marketing 国际贸易实务 Practice of InternationalTrade 服务业营销 Service Industry Marketing企业伦理 Enterprise Ethics 新产品开发 New Products Development管理信息系统 System of Management Information 运筹学 Operational Research 保险学 Insurance管理会计 Managerial Accounting 国际贸易 International Trade国际⾦融 International Finance证券投资学 Security Analysis and Investment 国际结算 International BalanceFinancial Marketing证券投资学Security Analysis and Investment 财务学专业财务报告分析Analysis of Financial Statement 国际⾦融International Finance保险学Insurance 财务案例分析Case Analysis of FinanceManagement 国际财务管理International Financial Management 资产评估Assets Appraisal项⽬评估Projects Appraisal 财务管理Financial Management 运筹学Operational Research 管理会计Managerial Accounting 管理学Principles of Management 统计学Principles of Statistics宏观经济学Macroeconomics管理信息系统Systems of Management Information 策略管理Strategic Management 微观经济学Microeconomics 微积分Calculus会计专业会计学Accounting Principles 管理会计Managerial Accounting 会计信息系统Accounting Information Systems 财务管理Financial Management 财务报告分析Analysis of Financial Statement 国际会计International Accounting成本会计Cost Accounting 审计学Auditing Principles 投资学Investment Principles 货币银⾏学Money and Banking国际⾦融International Finance 统计学Principle of Stat财税法规与税务会计Laws and Regulations of Financeand Taxes预算会计Budget Accounting 会计研究⽅法Accounting Research Methods 内部审计与政府审计Internal Auditing and GovernmentAuditing会计审计实务Accounting and Auditing Practice 经济计量学Economic Metrology会计职业道德与责任Accounting Ethics and Responsibilities国际会计专题International AccountingSpecial Subject 微观经济学Microeconomics 货币银⾏学Money and Banking。

各专业课程英文翻译(精心整理)

各专业课程英文翻译(精心整理)生物及医学专业课程汉英对照表应用生物学 Applied Biology 医学技术 Medical Technology细胞生物学 Cell Biology 医学 Medicine生物学 Biology 护理麻醉学 Nurse Anesthesia进化生物学 Evolutionary Biology 口腔外科学 Oral Surgery海洋生物学 Marine Biology 口腔/牙科科学 Oral/Dental Sciences微生物学 Microbiology 骨科医学 Osteopathic Medicine分子生物学 Molecular Biology 耳科学 Otology医学微生物学 Medical Microbiology 理疗学 Physical Therapy口腔生物学 Oral Biology 足病医学 Podiatric Medicine寄生物学 Parasutology 眼科学 Ophthalmology植物生物学 Plant Physiology 预防医学 Preventive Medicine心理生物学 Psychobiology 放射学 Radiology放射生物学 Radiation Biology 康复咨询学 Rehabilitation Counseling理论生物学 Theoretical Biology 康复护理学 Rehabilitation Nursing野生生物学 Wildlife Biology 外科护理学 Surgical Nursing环境生物学 Environmental Biology 治疗学 Therapeutics运动生物学 Exercise Physiology 畸形学 Teratology有机体生物学 Organismal Biology 兽医学 V eterinary Sciences生物统计学 Biometrics 牙科卫生学 Dental Sciences生物物理学 Biophysics 牙科科学 Dentistry生物心理学 Biopsychology 皮肤学 Dermatology生物统计学 Biostatistics 内分泌学 Endocrinology生物工艺学 Biotechnology 遗传学 Genetics生物化学 Biological Chemistry 解剖学 Anatomy生物工程学 Biological Engineering 麻醉学 Anesthesia生物数学 Biomathematics 临床科学 Clinical Science生物医学科学 Biomedical Science 临床心理学 Clinical Psychology细胞生物学和分子生物学 Celluar and Molecular Biology精神病护理学 Psychiatric Nursing力学专业数学分析 Mathematical Analysis 高等代数与几何 Advanced Algebra and Geometry 常微分方程 Ordinary Differential Equation 数学物理方法 Methods in Mathematical Physics 计算方法 Numerical Methods 理论力学 Theoretical Mechanics材料力学 Mechanics of Materials 弹性力学 Elasticity流体力学 Fluid Mechanics 力学实验 Experiments in Solid Mechanics机械制图 Machining Drawing 力学概论 Introduction to Mechanics气体力学 Gas Dynamics 计算流体力学 Computational Fluid Mechanics 弹性板理论 Theory of Elastic Plates 粘性流体力学 V iscous Fluid Flow弹性力学变分原理 V ariational Principles inElasticity 有限元法 Finite Element Method 塑性力学 Introduction of Plasticity经典力学中的数学方法 Mathematical Methods of ClassicalMechanics机器人动力学 Dynamics of Robots 自动控制原理 Principles of Automatic Control 优化计算与最化控制 Optimization and OptimalControl计算机图形学 Computer Graphics 概率与统计 Probability and Statistics专业英语 English for Mechanics 振动理论 Theory of V ibration程序设计方法(C和FORTRAN) Programming in C & FORTRAN水动力学 Hydrodynamics 计算机图象处理 Image Processing光测力学 Photo Mechanics 断裂力学 Fracture Mechanics高等动力学 Advanced Dynamics 摄动方法 Perturbation Methods机械设计与Auto CAD Machinery Designing and AutoCAD信息显示(可视化) V isualization微机原理 Principles of Personal Computer 复变函数 Complex Function企业管理专业管理学 Principles of Management 微观经济学 Microeconomics宏观经济学 Macroeconomics 管理信息系统 Systems of Management Information 产业经济学 Industrial Economics 财务管理 Financial Management项目评估 Projects Appraisal 战略管理 Strategic Management管理沟通 Management Negotiation 国际商务谈判 Negotiation on Business Affairs跨国公司专题研究 Special Researchof multinational corporation国际贸易 InternationalTrade 国际营销研究 International Marketing Research公司组织与管理 Organization and Managementof Corporate战略管理 Strategic Management 生产管理研究 Operation Management企业伦理 Enterprise Ethics 组织行为学 Organizational Behavior运筹学 Operational Research 人力资源管理 Human Resource Management信息管理专业高等数学 Higher Mathematics 信息存储与检索 Information Retrieval andStorage数据库系统 Database 信息服务与用户 information Service and UserStudy 信息管理概论 Introduction to InformationManagement信息经济学 Information Economics 企业信息化工程 Enterprise Informationalization社会实践 Practical Work 信息分析与决策Information Analysis andPolicy Making 管理学原理 Principles of Management 信息政策与法规 Information Policy and Law信息组织 Information Organization 计算机网络 Computer Networks管理信息系统 Management Information System 线性代数 Linear Algebra决策分析 Policy Making 离散数学 Discrete Mathematics概率统计 Statistics and ProbabilityTheory 生产与运作管理 Production Management 电子商务 Electronic Commerce 信息系统安全与保密 Information System Security 政府信息化工程 Government Informationalization广告实务 Practice of Advertisement 多媒体技术 Multimedia操作系统 Operating System 信息科学基础 Foundations of InformationScience 经济信息管理 Economic Information Management 专业英语 Specialty English微机基础 Principles of Microcomputers 文献计量学 Bibliometrics电子出版技术 Electronic Publishing 广告概论 Introduction to Advertisement信息环境论 Information Environments 传播学原理 Principles of CommunicationTheory 知识产权法学 Law of Intelligence Property 组织行为学 Studies of Organization货币银行学专业货币银行学 Money and Banking 管理信息系统 System of Management Information 宏观经济学 Macroeconomics 运筹学 Operational Research策略管理 Strategic Management 保险学 Insurance银行会计 Bank Accounting 管理会计 Managerial Accounting运筹学 Operational Research 国际贸易 International Trade财务管理 Financial Management 国际金融 International Finance租赁与信托 Hiring and Affiancing 证券投资学 Security Analysis and Investment商业银行实务 Practice of Business Bank 国际结算 International Balance项目评估 Projects Appraisal 金融市场学 Financial Marketing人力资源管理 Human Resource Management财务报告分析 Analysis of Financial Statement财务案例分析 Case Analysis of FinancialManagement物理专业热学 Thermodynamics 力学 Mechanics光学 Optics 电磁学 Electromagnetism计算概论 Computing Generality 普通物理实验 General Physics Laboratory固体磁性及应用基础 Magnetism of the Solid Stateand its Application衍射物理（固体结构分析） Diffraction Physics (Structureof Solid Analysis)科研实用软件 Utility Software for ScientificResearch计算机模拟方法 Computer Simulation Methods激光原理、技术与应用 The Principle, Techniqueand Application of Laser材料物理 Materials Physics 近代光学和光电子学 Modern Optics and Optoelectronics 现代固体物理 Modern Solid State Physics 粒子物理 Particle Physics物理宇宙学基础 Elements of Cosmology Physics 固体物理 Solid State Physics原子物理 Atomic Physics 量子力学 Quantum Mechanics理论力学 Theoretical Mechanics 电动力学 Electrodynamics普通物理综合实验 Synthetical Experiments ofGeneral Physics市场营销学专业营销管理 Marketing Management 公共关系 Public Relationship国际贸易 International Trade 消费者行为 Consumer Behavior管理信息系统 Systems of Management Information 营销调研 Marketing Research推销学 Sales Strategies 国际金融 International Finance营销预测与规划 Marketing Forecasting andPlanning销售渠道管理 ales Channels Management 管理学 Principles of Management国际市场营销 International Marketing 商业谈判 Business Negotiation广告管理 Advertising Management 营销案例分析 Case Studies of Marketing国际贸易实务 Practice of InternationalTrade 服务业营销 Service Industry Marketing企业伦理 Enterprise Ethics 新产品开发 New Products Development财务学专业货币银行学 Money and Banking 证券投资学 Security Analysis and Investment 财务报告分析 Analysis of Financial Statement 国际金融 International Finance保险学 Insurance 财务案例分析 Case Analysis of FinanceManagement 国际财务管理 International Financial Management 资产评估 Assets Appraisal项目评估 Projects Appraisal 宏观经济学 Macroeconomics财务管理 Financial Management 管理信息系统 Systems of Management Information 运筹学 Operational Research 策略管理 Strategic Management管理会计 Managerial Accounting 微观经济学 Microeconomics管理学 Principles of Management 微积分 Calculus统计学 Principles of Statistics会计专业会计学 Accounting Principles 成本会计 Cost Accounting管理会计 Managerial Accounting 审计学 Auditing Principles会计信息系统 Accounting Information Systems 投资学 Investment Principles财务管理 Financial Management 货币银行学 Money and Banking财务报告分析 Analysis of Financial Statement 国际金融 International Finance国际会计 International Accounting 统计学 Principle of Stat财税法规与税务会计 Laws and Regulations of Financeand Taxes预算会计 Budget Accounting 会计研究方法 Accounting Research Methods 内部审计与政府审计 Internal Auditing and GovernmentAuditing会计审计实务 Accounting and Auditing Practice 经济计量学 Economic Metrology会计职业道德与责任 Accounting Ethics and Responsibilities国际会计专题 International AccountingSpecial Subject 微观经济学 Microeconomics。

分子生物学英文文献6

Chapter19Detection and Quantitative Analysis of Small RNAs by PCR Seungil Ro and Wei YanAbstractIncreasing lines of evidence indicate that small non-coding RNAs including miRNAs,piRNAs,rasiRNAs, 21U endo-siRNAs,and snoRNAs are involved in many critical biological processes.Functional studies of these small RNAs require a simple,sensitive,and reliable method for detecting and quantifying levels of small RNAs.Here,we describe such a method that has been widely used for the validation of cloned small RNAs and also for quantitative analyses of small RNAs in both tissues and cells.Key words:Small RNAs,miRNAs,piRNAs,expression,PCR.1.IntroductionThe past several years have witnessed the surprising discovery ofnumerous non-coding small RNAs species encoded by genomesof virtually all species(1–6),which include microRNAs(miR-NAs)(7–10),piwi-interacting RNAs(piRNAs)(11–14),repeat-associated siRNAs(rasiRNAs)(15–18),21U endo-siRNAs(19),and small nucleolar RNAs(snoRNAs)(20).These small RNAsare involved in all aspects of cellular functions through direct orindirect interactions with genomic DNAs,RNAs,and proteins.Functional studies on these small RNAs are just beginning,andsome preliminaryﬁndings have suggested that they are involvedin regulating genome stability,epigenetic marking,transcription,translation,and protein functions(5,21–23).An easy and sensi-tive method to detect and quantify levels of these small RNAs inorgans or cells during developmental courses,or under different M.Sioud(ed.),RNA Therapeutics,Methods in Molecular Biology629,DOI10.1007/978-1-60761-657-3_19,©Springer Science+Business Media,LLC2010295296Ro and Yanphysiological and pathophysiological conditions,is essential forfunctional studies.Quantitative analyses of small RNAs appear tobe challenging because of their small sizes[∼20nucleotides(nt)for miRNAs,∼30nt for piRNAs,and60–200nt for snoRNAs].Northern blot analysis has been the standard method for detec-tion and quantitative analyses of RNAs.But it requires a relativelylarge amount of starting material(10–20μg of total RNA or>5μg of small RNA fraction).It is also a labor-intensive pro-cedure involving the use of polyacrylamide gel electrophoresis,electrotransfer,radioisotope-labeled probes,and autoradiogra-phy.We have developed a simple and reliable PCR-based methodfor detection and quantiﬁcation of all types of small non-codingRNAs.In this method,small RNA fractions are isolated and polyAtails are added to the3 ends by polyadenylation(Fig.19.1).Small RNA cDNAs(srcDNAs)are then generated by reverseFig.19.1.Overview of small RNA complementary DNA(srcDNA)library construction forPCR or qPCR analysis.Small RNAs are polyadenylated using a polyA polymerase.ThepolyA-tailed RNAs are reverse-transcribed using a primer miRTQ containing oligo dTsﬂanked by an adaptor sequence.RNAs are removed by RNase H from the srcDNA.ThesrcDNA is ready for PCR or qPCR to be carried out using a small RNA-speciﬁc primer(srSP)and a universal reverse primer,RTQ-UNIr.Quantitative Analysis of Small RNAs297transcription using a primer consisting of adaptor sequences atthe5 end and polyT at the3 end(miRTQ).Using the srcD-NAs,non-quantitative or quantitative PCR can then be per-formed using a small RNA-speciﬁc primer and the RTQ-UNIrprimer.This method has been utilized by investigators in numer-ous studies(18,24–38).Two recent technologies,454sequenc-ing and microarray(39,40)for high-throughput analyses of miR-NAs and other small RNAs,also need an independent method forvalidation.454sequencing,the next-generation sequencing tech-nology,allows virtually exhaustive sequencing of all small RNAspecies within a small RNA library.However,each of the clonednovel small RNAs needs to be validated by examining its expres-sion in organs or in cells.Microarray assays of miRNAs have beenavailable but only known or bioinformatically predicted miR-NAs are covered.Similar to mRNA microarray analyses,the up-or down-regulation of miRNA levels under different conditionsneeds to be further validated using conventional Northern blotanalyses or PCR-based methods like the one that we are describ-ing here.2.Materials2.1.Isolation of Small RNAs, Polyadenylation,and Puriﬁcation 1.mirVana miRNA Isolation Kit(Ambion).2.Phosphate-buffered saline(PBS)buffer.3.Poly(A)polymerase.4.mirVana Probe and Marker Kit(Ambion).2.2.Reverse Transcription,PCR, and Quantitative PCR 1.Superscript III First-Strand Synthesis System for RT-PCR(Invitrogen).2.miRTQ primers(Table19.1).3.AmpliTaq Gold PCR Master Mix for PCR.4.SYBR Green PCR Master Mix for qPCR.5.A miRNA-speciﬁc primer(e.g.,let-7a)and RTQ-UNIr(Table19.1).6.Agarose and100bp DNA ladder.3.Methods3.1.Isolation of Small RNAs 1.Harvest tissue(≤250mg)or cells in a1.7-mL tube with500μL of cold PBS.T a b l e 19.1O l i g o n u c l e o t i d e s u s e dN a m eS e q u e n c e (5 –3 )N o t eU s a g em i R T QC G A A T T C T A G A G C T C G A G G C A G G C G A C A T G G C T G G C T A G T T A A G C T T G G T A C C G A G C T A G T C C T T T T T T T T T T T T T T T T T T T T T T T T T V N ∗R N a s e f r e e ,H P L CR e v e r s e t r a n s c r i p t i o nR T Q -U N I r C G A A T T C T A G A G C T C G A G G C A G GR e g u l a r d e s a l t i n gP C R /q P C Rl e t -7a T G A G G T A G T A G G T T G T A T A G R e g u l a r d e s a l t i n gP C R /q P C R∗V =A ,C ,o r G ;N =A ,C ,G ,o r TQuantitative Analysis of Small RNAs299 2.Centrifuge at∼5,000rpm for2min at room temperature(RT).3.Remove PBS as much as possible.For cells,remove PBScarefully without breaking the pellet,leave∼100μL of PBS,and resuspend cells by tapping gently.4.Add300–600μL of lysis/binding buffer(10volumes pertissue mass)on ice.When you start with frozen tissue or cells,immediately add lysis/binding buffer(10volumes per tissue mass)on ice.5.Cut tissue into small pieces using scissors and grind it usinga homogenizer.For cells,skip this step.6.Vortex for40s to mix.7.Add one-tenth volume of miRNA homogenate additive onice and mix well by vortexing.8.Leave the mixture on ice for10min.For tissue,mix it every2min.9.Add an equal volume(330–660μL)of acid-phenol:chloroform.Be sure to withdraw from the bottom phase(the upper phase is an aqueous buffer).10.Mix thoroughly by inverting the tubes several times.11.Centrifuge at10,000rpm for5min at RT.12.Recover the aqueous phase carefully without disrupting thelower phase and transfer it to a fresh tube.13.Measure the volume using a scale(1g=∼1mL)andnote it.14.Add one-third volume of100%ethanol at RT to the recov-ered aqueous phase.15.Mix thoroughly by inverting the tubes several times.16.Transfer up to700μL of the mixture into aﬁlter cartridgewithin a collection bel theﬁlter as total RNA.When you have>700μL of the mixture,apply it in suc-cessive application to the sameﬁlter.17.Centrifuge at10,000rpm for15s at RT.18.Collect theﬁltrate(theﬂow-through).Save the cartridgefor total RNA isolation(go to Step24).19.Add two-third volume of100%ethanol at RT to theﬂow-through.20.Mix thoroughly by inverting the tubes several times.21.Transfer up to700μL of the mixture into a newﬁlterbel theﬁlter as small RNA.When you have >700μL of theﬁltrate mixture,apply it in successive appli-cation to the sameﬁlter.300Ro and Yan22.Centrifuge at10,000rpm for15s at RT.23.Discard theﬂow-through and repeat until all of theﬁltratemixture is passed through theﬁlter.Reuse the collectiontube for the following washing steps.24.Apply700μL of miRNA wash solution1(working solu-tion mixed with ethanol)to theﬁlter.25.Centrifuge at10,000rpm for15s at RT.26.Discard theﬂow-through.27.Apply500μL of miRNA wash solution2/3(working solu-tion mixed with ethanol)to theﬁlter.28.Centrifuge at10,000rpm for15s at RT.29.Discard theﬂow-through and repeat Step27.30.Centrifuge at12,000rpm for1min at RT.31.Transfer theﬁlter cartridge to a new collection tube.32.Apply100μL of pre-heated(95◦C)elution solution orRNase-free water to the center of theﬁlter and close thecap.Aliquot a desired amount of elution solution intoa1.7-mL tube and heat it on a heat block at95◦C for∼15min.Open the cap carefully because it might splashdue to pressure buildup.33.Leave theﬁlter tube alone for1min at RT.34.Centrifuge at12,000rpm for1min at RT.35.Measure total RNA and small RNA concentrations usingNanoDrop or another spectrophotometer.36.Store it at–80◦C until used.3.2.Polyadenylation1.Set up a reaction mixture with a total volume of50μL in a0.5-mL tube containing0.1–2μg of small RNAs,10μL of5×E-PAP buffer,5μL of25mM MnCl2,5μL of10mMATP,1μL(2U)of Escherichia coli poly(A)polymerase I,and RNase-free water(up to50μL).When you have a lowconcentration of small RNAs,increase the total volume;5×E-PAP buffer,25mM MnCl2,and10mM ATP should beincreased accordingly.2.Mix well and spin the tube brieﬂy.3.Incubate for1h at37◦C.3.3.Puriﬁcation 1.Add an equal volume(50μL)of acid-phenol:chloroformto the polyadenylation reaction mixture.When you have>50μL of the mixture,increase acid-phenol:chloroformaccordingly.2.Mix thoroughly by tapping the tube.Quantitative Analysis of Small RNAs3013.Centrifuge at10,000rpm for5min at RT.4.Recover the aqueous phase carefully without disrupting thelower phase and transfer it to a fresh tube.5.Add12volumes(600μL)of binding/washing buffer tothe aqueous phase.When you have>50μL of the aqueous phase,increase binding/washing buffer accordingly.6.Transfer up to460μL of the mixture into a puriﬁcationcartridge within a collection tube.7.Centrifuge at10,000rpm for15s at RT.8.Discard theﬁltrate(theﬂow-through)and repeat until allof the mixture is passed through the cartridge.Reuse the collection tube.9.Apply300μL of binding/washing buffer to the cartridge.10.Centrifuge at12,000rpm for1min at RT.11.Transfer the cartridge to a new collection tube.12.Apply25μL of pre-heated(95◦C)elution solution to thecenter of theﬁlter and close the cap.Aliquot a desired amount of elution solution into a1.7-mL tube and heat it on a heat block at95◦C for∼15min.Open the cap care-fully because it might be splash due to pressure buildup.13.Let theﬁlter tube stand for1min at RT.14.Centrifuge at12,000rpm for1min at RT.15.Repeat Steps12–14with a second aliquot of25μL ofpre-heated(95◦C)elution solution.16.Measure polyadenylated(tailed)RNA concentration usingNanoDrop or another spectrophotometer.17.Store it at–80◦C until used.After polyadenylation,RNAconcentration should increase up to5–10times of the start-ing concentration.3.4.Reverse Transcription 1.Mix2μg of tailed RNAs,1μL(1μg)of miRTQ,andRNase-free water(up to21μL)in a PCR tube.2.Incubate for10min at65◦C and for5min at4◦C.3.Add1μL of10mM dNTP mix,1μL of RNaseOUT,4μLof10×RT buffer,4μL of0.1M DTT,8μL of25mM MgCl2,and1μL of SuperScript III reverse transcriptase to the mixture.When you have a low concentration of lig-ated RNAs,increase the total volume;10×RT buffer,0.1M DTT,and25mM MgCl2should be increased accordingly.4.Mix well and spin the tube brieﬂy.5.Incubate for60min at50◦C and for5min at85◦C toinactivate the reaction.302Ro and Yan6.Add1μL of RNase H to the mixture.7.Incubate for20min at37◦C.8.Add60μL of nuclease-free water.3.5.PCR and qPCR 1.Set up a reaction mixture with a total volume of25μL ina PCR tube containing1μL of small RNA cDNAs(srcD-NAs),1μL(5pmol of a miRNA-speciﬁc primer(srSP),1μL(5pmol)of RTQ-UNIr,12.5μL of AmpliTaq GoldPCR Master Mix,and9.5μL of nuclease-free water.ForqPCR,use SYBR Green PCR Master Mix instead of Ampli-Taq Gold PCR Master Mix.2.Mix well and spin the tube brieﬂy.3.Start PCR or qPCR with the conditions:95◦C for10minand then40cycles at95◦C for15s,at48◦C for30s and at60◦C for1min.4.Adjust annealing Tm according to the Tm of your primer5.Run2μL of the PCR or qPCR products along with a100bpDNA ladder on a2%agarose gel.∼PCR products should be∼120–200bp depending on the small RNA species(e.g.,∼120–130bp for miRNAs and piRNAs).4.Notes1.This PCR method can be used for quantitative PCR(qPCR)or semi-quantitative PCR(semi-qPCR)on small RNAs suchas miRNAs,piRNAs,snoRNAs,small interfering RNAs(siRNAs),transfer RNAs(tRNAs),and ribosomal RNAs(rRNAs)(18,24–38).2.Design miRNA-speciﬁc primers to contain only the“coresequence”since our cloning method uses two degeneratenucleotides(VN)at the3 end to make small RNA cDNAs(srcDNAs)(see let-7a,Table19.1).3.For qPCR analysis,two miRNAs and a piRNA were quan-titated using the SYBR Green PCR Master Mix(41).Cyclethreshold(Ct)is the cycle number at which theﬂuorescencesignal reaches the threshold level above the background.ACt value for each miRNA tested was automatically calculatedby setting the threshold level to be0.1–0.3with auto base-line.All Ct values depend on the abundance of target miR-NAs.For example,average Ct values for let-7isoforms rangefrom17to20when25ng of each srcDNA sample from themultiple tissues was used(see(41).Quantitative Analysis of Small RNAs3034.This method ampliﬁes over a broad dynamic range up to10orders of magnitude and has excellent sensitivity capable ofdetecting as little as0.001ng of the srcDNA in qPCR assays.5.For qPCR,each small RNA-speciﬁc primer should be testedalong with a known control primer(e.g.,let-7a)for PCRefﬁciency.Good efﬁciencies range from90%to110%calcu-lated from slopes between–3.1and–3.6.6.On an agarose gel,mature miRNAs and precursor miRNAs(pre-miRNAs)can be differentiated by their size.PCR prod-ucts containing miRNAs will be∼120bp long in size whileproducts containing pre-miRNAs will be∼170bp long.However,our PCR method preferentially ampliﬁes maturemiRNAs(see Results and Discussion in(41)).We testedour PCR method to quantify over100miRNAs,but neverdetected pre-miRNAs(18,29–31,38). 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生物专业文献翻译技巧

生物专业文献翻译技巧乔凌燕刘勇(阜阳师范学院安徽阜阳236037)【摘要】在高等院校生命科学类专业选修课程之一生物专业英语中,翻译结构复杂的句子是生物专业英语教学中的困难点。

本文将以生物专业英语教材中的具体语句作为例子,阐述了中长复杂语句的名词性短语、被动时态、用It 引导的强调句和主语从句的转换翻译技巧,与倒置法、拆句法等在翻译过程与生物专业的巧妙结合。

【关键词】生物专业英语;翻译技巧;拆句法;倒置法【中图分类号】H319【文献标识码】B 【文章编号】2095-3089(2016)11-0040-01生物专业英语是在高等院校中由生命科学专业开展的一门专业选修课程,通常在学生学习完基础英语课程和相关专业课程后开展的。

该课程是以使学生能轻松阅读国外相关的专业文献和了解国内外本专业发展的最新动态,能使用英语进行专业学术交流,并拥有能检索专业英语文献和撰写英语专业论文能力为教学目标。

生物专业,是描述自然界生物的特点和发展过程等。

同时生物专业英语表达要求客观精确,句子的结构严密。

生物专业英语通常具有词汇的词形较长,被动句出现频率高、专业高、词性转换频繁强等特点,较为枯燥乏味。

在教学中,发现学生主要是被专业文献中的难句和长句翻译所难住。

因此,难句和长句的翻译是教师提高生物专业英语教学质量的重要突破点与难点,也是提高学生实际阅读生物英语参考文献能力的关键所在。

一、以it 引导的强调句或主语从句的翻译it 引导的主语从句和强调句在生物英语专业中是非常常见的。

通常情况,强调句型是为了强调某一成分,主语从句则是为了保证句子结构的平衡。

进行这种句子的翻译要注意汉语的语法规则与逻辑顺序:有it 引导的主语从句只用译出句子中的逻辑主语意思即可;由it 引导的强调句,主要是为了强调,需在译文中加上相应强调词,如“就是’“正是”等。

二、将被动转换成主动生物专业英语作为科技英语的一种,通常是表达事理,常不用说出当事人;而且,常为了表示客观,通常会避免使用具有较强的主观性的主动时态,而是尽量使用被动语态。

分子生物学(英文版)

Chapter 3 Nucleic Acid1. Physical and chemical structure of DNA●Double-stranded helix● Major groove and minor groove● Base pairing● The two strands are antiparallel● G+C content (percent G+C)● Satellite DNASatellite DNA consists of highly repetitive DNA and is so called because repetitions of a short DNA sequence tend to produce a different frequency of the nucleotides adenine, cytosine， guanine and thymine， and thus have a different density from bulk DNA — such that they form a second or ’satellite’ band when genomic DNA is separated on a density gradient。

2。

Alternate DNA structureTwo bases have been extruded from base stacking at the junction. The white line goes from phosphate to phosphate along the chain。

O is shown red, N blue， P yellow and C grey.3. Circular and superhelical DNADNA can also form a double-stranded, covalently-closed circle。

分子生物学常用参考书目

二十一世纪是分子生物学发展的世纪，生命科学将进入一个新的时代——后基因组时代postgenomics
二十一世纪分子生物学发展的趋势：
1.功能基因组学 functional genomics 依附于对DNA序列的了解，应用基因组学的知识和工具
去了解影响发育和整个生物体的特征序列表达谱。酿酒酵母16条染色体的全部序列于1996年完成。
…
1997
Wilmut成功获得克隆羊—Dolly诞生；
1998
Renard 克隆牛诞生（体细胞→个体）；
…
2000 ,6.26 中、美、日、德、法、英6国，宣布人类基因组草图发表。
2000 ,10月科学家宣布将于2001年3月完成河豚鱼的基因组测序。
2000,12月14日英美等国科学家宣布绘出拟南芥基因组的完整图谱。
2003年4月14日六国科学家完成了人类基因组序列图的绘制，实现了人类基因组计划的所有目标。
二十世纪是以核酸为研究核心，带动分子生物学向纵深发展：
•
50年代双螺旋结构
•
60年代操纵子学说
•
70年代DNA重组
•
80年代PCR技术
•
90年代DNA测序
生命科学从宏观→微观→宏观；由分析→综合的时代。
分子生物学常用参考书目
2024/2/1
第一章绪论
一、什么是分子生物学？
Instant Notes in Molecular Biology
---Turner et al.
Molecular biology seeks to explain the relationships between the structure and function of biological molecules and how these relationships contribute to the operation and control of biochemical processes.

第8讲翻译（2）

2）43S小亚基前起始复合物
含eIF3 需要GTP
3）小亚基起始复合物
43S起始前复合体识别5’帽结构，并通过eIF3与eIF4F间的相互作用募集43S 起始前复合体到mRNA上
Cap-dependent小亚基起始复合物的形成
形成多种起始复合物需要起始因子及一个特殊的起始tRNA；
•核糖体小亚基负责对模板mRNA进行序列特异性识别， •大亚基负责携带氨基酸及tRNA的功能，肽键的形成、AAtRNA、肽基-tRNA的结合等主要在大亚基上。
核糖体上重要位点
去氨酰-tRNA 肽基酰-tRNA 氨酰-tRNA
• A位点(aminoacyl site),新到来的氨酰-tRNA的结合位点; • P位点(peptidyl site),肽酰-tRNA结合位点; • E位点(Exit site), 释放去氨酰-tRNA的位点。 • 只有(f)Met-tRNAfMet能与第一个P位点相结合，其它所有 tRNA都必须通过A位点到达P位点，再由E位点离开核糖体。
mRNA进入和离开的通道位于小亚基
E
P
A
mRNA
14 Angstroms
Location of tRNAs
Cate et al., Science 1999
• 每一个tRNA结合位点都横跨核糖体的两个亚基，位于大、小亚基的交界面。
Channel for polypeptide chain exiting locates in the large subunit
A: Without mRNA. B: With capped and
polyadenylated luciferase mRNA. C–E: Higher magnification of c, d

分子生物学翻译

分子生物学翻译
• 翻译: 是蛋白质生物合成过程中的第一步。翻译是根据遗传密码的中心法则，将成熟的mRNA分子中碱基的排列顺序（核苷酸序列）解码，并生成对应的特定氨基酸序列的过程。但也有许多转录生成的RNA，如 tRNA、rRNA和snRNA等并不被翻译为氨基酸序列。
• 翻译分作三个阶段：起始、延长、终止。
NH2
fMet
Cys
His
Ala
tRNA
mRNA 5’
Direction of translation
GU ACG U A UGCA UGC AUG C U ACGUU A
3’
图 6.2 翻译时RNA不能直接作为生产氨基酸模板
• 遗传密码是一组规则，将DNA或RNA序列以三个核苷酸为一组的密码子翻译为蛋白质的氨基酸序列，以用于蛋白质合成。几乎所有的生物都使用同样的遗传密码，称为标准遗传密码；即使是非细胞结构的病毒，它们也是使用标准遗传密码。但是也有少数生物使用一些稍微不同的遗传密码。
(L1 ~ L50)
S1 S2 S3
S33
33 proteins of small subunit
(S1 ~ S33)
60S subunit 40S subunit
图 6.4 原核生物与真核生物核糖体的组成
70S prokaryotic ribosome
80S eukaryotic ribosome
6.2.1 起始
• 多聚核糖体：原核生物中带有很多核糖体的mRNA称为多聚核糖体。
• 开放阅读框（Open Reading Frame）就是直接翻译成蛋白质的那段DNA序列。从atg开始到终止密码子结束，中间没有内含子。
• 非翻译区（UnTranslated Regions) 转录产物开头和末尾不翻译成蛋白质的那段序列。

分子生物学翻译

fMet fMet
Tu GTP
5'
AUG
3'
进位
成肽
转位
（四）真核生物延长过程
真核生物肽链合成的延长过程与原核基本相似，但有不同的反应体系和延长因子。
另外，真核细胞核蛋白体没有E位，转位时卸载的tRNA直接从P位脱落。
三、肽链合成的终止
当mRNA上终止密码出现后，多肽链合成停止，肽链从肽酰-tRNA中释出， mRNA、核蛋白体等分离，这些过程称为肽链合成终止。
胞浆胞浆
tRNA rRNA
74-95个核苷酸
28S，5400个核苷酸 18S，2100个核苷酸 5.8S，160个核苷酸 5S， 120个核苷酸
转运氨基酸与密码子识别
构成核糖体，蛋白质合成场所
S：沉降系数（1S=10-13秒）
碱基数量：bp, Kb, Mb
原核生物16S rRNA的二级结构
（一）原核生物翻译起始复合物形成
• 核蛋白体大小亚基分离； • mRNA在小亚基定位结合； • 起始氨基酰-tRNA的结合； • 核蛋白体大亚基结合。
1. 核蛋白体大小亚基分离
IF-1 IF-3
2. mRNA在小亚基定位结合
5'
AUG
3'
IF-1
IF-3
S-D序列：
在原核生物mRNA起始密码AUG上游，存在4~9个富含嘌呤碱的一致性序列，如-AGGAGG-，称为S-D序列。又称为核蛋白体结合位点（ribosomal binding site，RBS)
氨基酸的活化形式：氨基酰－tRNA 氨基酸的活化部位：α－羧基氨基酸与tRNA连接方式：酯键氨基酸活化耗能：2个~P

cellular and molecular life sciences 参考文献格式

cellular and molecular life sciences参考文献格式标题：细胞与分子生命科学的综述引言概述：细胞与分子生命科学是一门研究生物体内细胞和分子层面的学科，涉及细胞结构、功能、分子生物学、遗传学等多个方面。

本文将从细胞结构、功能、分子生物学、遗传学和生物技术五个大点出发，详细阐述细胞与分子生命科学的相关内容。

正文内容：1. 细胞结构1.1 细胞膜结构：细胞膜是细胞的外层，由磷脂双分子层和蛋白质组成。

细胞膜的结构和功能是细胞内外物质交换的关键。

1.2 细胞器结构：细胞器是细胞内的功能区域，包括核、线粒体、内质网等。

每个细胞器都有特定的结构和功能，协同工作维持细胞的正常运作。

2. 细胞功能2.1 细胞分裂：细胞分裂是细胞生命周期中的重要过程，包括有丝分裂和无丝分裂。

细胞分裂的调控对于生物体的生长和发育至关重要。

2.2 细胞信号传导：细胞通过信号分子进行信息传递，调控细胞的生理功能。

细胞信号传导的研究有助于揭示细胞内的调控机制。

2.3 细胞凋亡：细胞凋亡是细胞自我调控的一种方式，通过程序性死亡维持组织和器官的平衡。

细胞凋亡的异常与多种疾病的发生密切相关。

3. 分子生物学3.1 DNA结构和功能：DNA是遗传信息的携带者，其双螺旋结构和碱基配对规律对于遗传信息的传递和复制具有重要意义。

3.2 RNA转录和翻译：RNA是DNA的转录产物，在蛋白质合成中起到重要作用。

RNA转录和翻译是基因表达的关键过程。

3.3 蛋白质结构和功能：蛋白质是生物体内功能最为多样的分子，其结构和功能的研究对于理解细胞活动和生物学过程具有重要意义。

4. 遗传学4.1 遗传物质的传递：遗传物质通过遗传信息的传递维持物种的遗传稳定性。

遗传物质的传递方式包括有性和无性两种。

4.2 基因调控：基因调控是遗传物质在细胞内的表达和调节过程，包括转录因子、表观遗传学等多个层面。

4.3 基因突变：基因突变是遗传信息发生变化的结果，对个体的遗传特征和疾病的发生有重要影响。

分子生物学名词解释英文

1.DNA Denaturation(变性) When duplex DNA molecules are subjected to conditions of pH ,temperature,or ionic strength that disrupt base-paring interactions, the DNA molecule has lost its’native conformation, and double helix DNA is separated to single strand DNA as individual randome coils.That is, the DNA is denatured.2.Renaturation（复性）Removing the denaturation factors slowly or in proper conditions, the denaturedDNA (ssDNA) restore native structure (dsDNA) and functions. This process is dependent on both DNA concentration and time.3.Hybridization （核酸分子杂交）when heterogeneous DNA or RNA are put together, they will become toheteroduplex via the base-pairing rules during renaturation if they are complementary in parts (not completely). This is called molecular hybridization.4.Hyperchromic effect （增色效应）The absorbance at 260 nm of a DNA solution increases when thedouble helix is separated into single strands because of the bases unstack.5.Ribozyme （核酶）are the RNA molecules with catalytic activity. The activity of these ribozymes ofteninvolves the cleavage of a nucleic acid.6.De novo synthesis (从头合成)De novo synthesis of nucleotides begins with their metabolic precursors:amino acids, ribose-5-phosphate, one carbon units, CO2. mostly in liver.7.Salvage pathways （补救合成）Salvage pathways recycle the free bases and nucleosides released fromnucleic acid breakdown. Mostly in brain and marrow.8.Semi-conservative replication (半保留复制）DNA is synthesized by separation of the strands of aparental duplex, each then acting as a template for synthesis of a complementary strand based on the base-paring rule. Each daughter molecule has one parental strand and one newly synthesized strand. 9.Telomere（端粒）：Specialized structure at the end of a linear eukaryotic chromosome, which consists ofproteins and DNA, tandem repeats of a short G-rich sequence on the 3 ' ending strand and its complementary sequence on the 5' ending strand, allows replication of the extreme 5' ends of the DNAwithout loss of genetic information and maintains the stability of eukaryote chromosome.10.Telomerase（端粒酶）An RNA-containing reverse transcriptase that using the RNA as a template, addsnucleotides to the 3 ' ending strand and thus prevents progressive shortening of eukaryotic linear DNA molecules during replication.11.Reverse transcription (逆转录）Synthesis of a double-strand DNA from an RNA template.12.Reverse transcriptase (逆转录酶）A DNA polymerase that uses RNA as its template.activity: RNA-dependent DNA polymerase; RNAse H;DNA-dependent DNA polymerase13.The central dogma (中心法则）It described that the flow of genetic information is from DNA to RNA andthen to protein. According to the central dogma, DNA directs the synthesis of RNA, and RNA then directs the synthesis of proteins.14.asymmetric transcription（不对称转录）1..Transcription generally involves only short segments of aDNA molecule, and within those segments only one of the two DNA strands serves as a template.2.The template strand of different genes is not always on the same strand of DNA. That is, in anychromosome, different genes may use different strands as template.15.template strand （模板链）The DNA strand that serves as a template for transcription. (The relationshipbetween template and transcript is base paring and anti-parallel)16.non-template strand (or coding strand)（编码连）The DNA strand that opposites to the templatestrand.(Note that it has the same sequence as the synthesized RNA, except for the replacement of U with T )17.promoter i s the DNA sequence at which RNA polymerase binds to initiate transcription. It is alwayslocated on the upstream of a gene.18.Split genes (断裂基因)Split genes are those in which regions that are represented in mature mRNAs orstructural RNAs (exons) are separated by regions that are transcribed along with exons in the primary RNA products of genes, but are removed from within the primary RNA molecule during RNA processingsteps (introns).19.Exon(外显子) can be expressed in primary transcript and are the sequences that are represented inmature RNA molecules, it encompasses not only protein-coding genes but also the genes for various RNA (such as tRNAs or rRNAs)20.Intron（内含子）can be expressed and be the intervening nucleotide sequences that are removed fromthe primary transcript when it is processed into a mature RNA.21.Spliceosome（剪切体）A multicomponent complex contains proteins and snRNAs that are involved inmRNA splicing.22.Translation（翻译）The process of protein synthesis in which the genetic information present in anmRNA molecule (transcribed from DNA) determines the sequence of amino acids by the genetic codons.Translation occurs on ribosomes.23.genetic codon（密码子）The genetic code is a triplet code read continuously from a fixed starting pointin each mRNA, also called triplet. Genetic code defines the relationship between the base sequence of mRNA and the amino acid sequence of polypeptide.24.Degeneracy of code（密码子简并性）One codon encodes only one amino acid;More than 2 codons can encode the same amino acid;Most codons that encode the same amino acid have the difference in the third base of the codon.25.ORF（开放阅读框架）The nucleotideacids sequences in mRNA molecule from 5’AUG to 3’stop codon(UAA UAG UGA). It consists of a group of contiguous nonoverlapping genetic codons encoding a whole protein. Usually, it includes more than 500 genetic codons.26.Shine-Dalgarno sequence（SD）is a sequence upstream the start codon in prokaryotic mRNA that canbase pairs to a •UCCU•sequence at or very near the 3' end of 16S rRNA, thereby binding the mRNA and small ribosomal subunit by each other.27.Polyribosome（多聚核糖体）Ribosomes(10~100) are tandemly arranged on one mRNA and move in thedirection of 5’to 3’.Such a complex of one mRNA and a number ofribosomes is called polyribosome.28.signal peptide（信号肽）It is a short conservative amino terminal sequence (13~36AA) that exists ona newly synthesized secretory protein. It can direct this protein to a specific locationwithin the cell. It is subsequently cleaved away by signal peptidase; also called signal sequence and targeting sequence.29.Operon（操纵子）: Bacteria have a simple general mechanism for coordinating the regulation of geneswhose products are involved in related processes: the genes are clustered on the chromosome and transcribed together. Most prokaryotic mRNAs are polycistronic. The single promoter requi red to initiate transcription of the cluster is the point where expression of all of the genes is regulated. The gene cluster, the promoter, and additional sequences that function in regulation are together called an operon. Operons that include 2 to 6 genes transcribed as a unit are common; some operons contain 20 or more genes.30.Housekeeping gene（管家基因）Genes that are expressed at a fairly consistent level throughout the cellcycle and from tissue to tissue. Usually involved in routine cellular metabolism. Often used for comparison when studying expression of other genes of interest.31.Trans-acting factors（反式作用因子）：Usually considered to be proteins, that bind to the cis-actingsequences to control gene expression. The properties of different trans-acting factors:subunits of RNA polymerasebind to RNA Polymerase to stabilize the initiation complexbind to all promoters at specific sequences but not to RNA Polymerase (TFIID factor which binds to the TATA box)bind to a few promoters and are required for transcription initiation32.Cis-acting elements（顺式作用元件）：DNA sequences in the vicinity of the structural portion of a genethat are required for gene expression. The properties of different cis-acting elements:contain short consensus sequencesmodules are related but not identicalnot fixed in location but usually within 200 bp upstream of the transcription start sitea single element is usually sufficient to confer a regulatory responsecan be located in a promoter or an enhancerassumed that a specific protein binds to the element and the presence of that protein is developmentally regulated33.Southern blotting：Genomic DNA (from tissues or cells) are cut by RE, separated by gelelectrophoresis and denatured in solution, then transferred to a nitrocellulose membrane for detecting specific DNA sequence by hybridization to a labeled probe. It can be used to quantitative and qualitative analyze genomic DNA, or analyze the recombinant plasmid and bacteriophage (screening DNA library).34.Northern blotting: RNA samples (from tissues or cells) are separated by gel electrophoresis anddenatured in solution, then transferred to a nitrocellulose membrane for detecting specific sequence by hybridization to a labeled probe. It can be used to detect the level of specific mRNA in some tissues (cells) and to compare the level of same gene expression in different tissues (cells) or at different development period.35.Western blotting：rotein samples are separated by PAGE electrophoresis, then electro-transferred to NCmembrane. The proteins on NC membrane hybridize with a specific antibody (1st antibody ), then the target protein binding with antibody is detected with a labeled secondary antibody (2nd antibody).Also called immunoblotting. It can be used to detect the specific protein, semi-quantify specific protein, etc.36.PBlotting technique（印迹）:Transfer (blot) biological macromolecules separated in the gel and fix themto nitrocellulose/nylon membrane by diffusion, electro-transferring or vacuum absorption, then detectit.37.Nucleic acid probe（探针）:DNA or RNA fragment labeled with radioisotope, biotin orfluorescent, is used to detect specific nucleic acid sequences by hybridization38.PCR: PCR is a technique for amplifying a specific DNA segment in vitro. The reaction system includeDNA template, T aq DNA pol, dNTP，short oligonucleotide primers, buffer containing Mg2+. The process including 3 steps: denature, annealing, extension39.DNA coloning（克隆）:T o clone a piece of DNA, DNA is cut into fragments using restriction enzymes. Thefragments are pasted into vectors that have been cut by the same restriction enzyme to form recombinant DNA. The recombinant DNA are needed to transfer and maintain DNA in a host cell. This serial process and related technique are called DNA coloning or genetic engineering.40.Genomic DNA library(基因组DNA文库) A genomic library is a set of clones that together representsthe entire genome of a given organism. The number of clones that constitute a genomic library depends on (1) the size of the genome in question and (2) the insert size tolerated by the particular cloning vector system. For most practical purposes, the tissue source of the genomic DNA is unimportant because each cell of the body contains virtually identical DNA (with some exceptions).41.cDNA library（cDNA文库）:A cDNA library represents a sample of the mRNA purified from a particularsource (either a collection of cells, a particular tissue, or an entire organism), which has been converted back to a DNA template by the use of the enzyme reverse transcriptase. It thus represents the genes that were being actively transcribed in that particular source under the physiological, developmental, or environmental conditions that existed when the mRNA was purified.42.α-complementation（α互补）:Some plasmid vectors such as pUC19 carry the alpha fragment of the lacZ gene. The alpha fragment is the amino-terminus of the beta-galactosidase. Typically, the mutant E. coli host strain only carry the omega fragment, which is the carboxy-terminus of the protein. Either omegaor alpha fragment alone is nonfunctional. When the vector containing lac Z introduced into mutant E.coli, both the alpha and omega fragments are present there is an interaction and a functionally intact beta-galactosidase protein can be produced. This interaction is called alpha complementation.43.Secondary messenger(第二信使) are some small signal molecules that are generated in the cell inresponse to extracellular signals. They can activate many other downstream components. The most important second messengers are: Ca2+, cAMP, cGMP, DAG, IP3, Cer, AA and its derivatives, etc.44.Adaptor protein（衔接蛋白）A specialized protein that links protein components of the signalingpathway, These proteins tend to lack any intrinsic enzymatic activity themselves but instead mediate specific protein-protein interaction that drive the formation of protein complexes.45.Scaffolding protein（支架蛋白）A protein that assembles interacting signaling proteins intomultimolecular, it recruits downstream effectors in a pathway and enhances specificity of the signal. 46.Oncogene（癌基因）A gene whose product is involved either in transforming cells in culture or ininducing cancer in animals including virus oncogene（v-onc）and cellular-oncogene（c-onc ）。

分子生物学英文文献

Mobile Genetic Elements 2:6, 267-271; November/December, 2012; © 2012 Landes BioscienceLETTER TO THE EDITORLETTER TO THE EDITORGAA repeats were shown to be the most unstable trinucleotide repeats in the pri-mates genome evolution by comparison of orthologous human and chimp loci.2 The instability of the GAA repeat in the first intron of the frataxin gene X25 is particu-larly well studied since it causes an inher-ited disorder, Friedreich ataxia (FRDA).3-6 I n Friedreich ataxia, once the length of the GAA repeat inside the frataxin gene (FXN GAA) reaches a certain threshold, the combined probability of its expan-sions and deletions in progeny of affected parents is about 85%.7 Deletions and con-tractions of the repeat in intergenerational transmissions can reach hundreds of base pairs.7 However, the FXN GAA repeat is much more stable in somatic cells.8 I t is relatively stable in blood, but shows some instability in dorsal root ganglia,9 which is responsible for some of the neurodegen-erative symptoms of Friedreich ataxia.5 GAA repeats were shown to be stable in FRDA fibroblasts cell lines and neuronal stem cells.10The question why the FXN GAA repeat is so much more stable in somatic cells than in intergenerational transmis-sions remains open. Recent studies in FRDA iPSCs that are closer to embryonic cells than somatic cells models, showed expansions of the GAA repeat with 100% probability.10,11 It is intriguing that all cells Complexes between two GAA repeats within DNA introduced into Cos-1 cellsMaria M. KrasilnikovaPennsylvania State University; University Park, PA USAKeywords: replication, GAA repeat, Friedreich ataxia, genome instability, chromatinCorrespondence to: Maria M. Krasilnikova; Email: muk19@ Submitted: 10/08/12; Revised: 12/09/12; Accepted: 12/10/12/10.4161/mge.23194in the iPSC cell lines that were analyzed were synchronously adding about two GAA repeats in each replication.The studies focused on the FXN GAA repeat provided many valuable insights; however, human genome contains many other GAA repeats: the human X chro-mosome, for instance, contains 44 GAA stretches with more than 100 repeats in each. About 30 GAA repeats were detected on the chromosome 4.12 GAA repeats mostly originated from the 3' end of the poly A associated with Alu elements.13I t is not known what makes repeats with the GAA motif most unstable com-pared with other trinucleotide repeats. It is possible that GAA repeats instability is caused by their ability to form non-B DNA structures. In vitro, GAA repeats can form triplexes,14,15 and sticky DNA structures.16 At the same time, hairpins 17 and paral-lel duplexes 18 have also been observed. When transcription is going through a GAA repeat, it can also form an R-loop, a DNA-RNA complex that leaves one of the complementary strands single-stranded.19 However, it is unclear whether these struc-tures indeed form in mammalian cells. If we assume that the instability of the GAA repeat is indeed associated with the struc-ture formation, it is still unclear why the structures would form in early embryo-genesis when the GAA expansion event in Friedriech ataxia is believed to occur,7 and do not form in somatic cells where the GAA repeat was shown to be more stable. I n our recent study, we hypothesize that the differences in chromatin structure are at least partially responsible for the differ-ences in the GAA repeats stability.1The propensity of GAA repeat to form a triplex structure may strongly depend on the structure of chromatin at the repeat and surrounding area.1 Consistent with other studies, we observed that formation of chromatin at an SV40-based plasmid introduced into mammalian cells occurs gradually: 8 h after transfection there are only occasional nucleosomes at the plas-mid, while by 72 h the nucleosome struc-ture is already regular.20 Our analysis of replication stalling at the repeat revealed that the repeat affects replication only in the first replication cycle, when chromatin is still at the formation stage. We believe that replication stalling at GAA is caused by a triplex structure that the GAA repeat adopts during transfection or inside the cell. In the subsequent replication cycles, replication was completely unaffected by the presence of the repeat, which is likely to be due to the inhibition of triplex for-mation by tight chromatin packaging.1Here we show the data that strengthen our previous observations and extend it to one more structure: a complex betweenWe have recently shown that GAA repeats severely impede replication elongation during the first replication cycle of transfected DNA wherein the chromatin is still at the formation stage.1 Here we extend this study by showing that two GAA repeats located within the same plasmid in the direct orientation can form complexes upon transient transfection of mammalian Cos-1 cells. However, these complexes do not form in DNA that went through several replication rounds in mammalian cells. We suggest that formation of such complexes in mammalian genomes can contribute to genomic instability.We studied replication of several SV40-based plasmids that contained two (GAA)57 repeats located at different posi-tions in Cos-1 cells (Figs. 1–3). I n each case, the cells were transiently transfected with 1 μg of each plasmid, and replication intermediates were isolated after about 30 h. This allowed us to observe replica-tion arcs that resulted from the plasmids that replicated for more than two rounds. However, some residual amount of the first replication cycle arcs can also be registered. Replication stalling at GAA repeats only occurs during the first repli-cation cycle of an SV40-based plasmid,1 hence we did not observe it in our system.For each of the plasmids, and for all patterns of restriction digests that we studied, we observed complexes between the two (GAA)57 repeats (indicated by red arrows in each of the figures). The migration of those complexes was differ-ent depending on the digest pattern, so the complexes were at different positions in 2D gel patterns, in agreement with our expectations based on their shapes.We did not observe replication stalling associated with these complexes. When this complex is formed, it migrates signifi-cantly slower on 2D gels; the replication of plasmids that contain such complexes should result in an extra replication arc originating from the complex position. However, the number of molecules that form this complex is significantly lower than the overall number of plasmids, and there may be not enough material to observe their replication.For the situation when the two GAA repeats were located in two different frag-ments upon PvuI digest (Fig. 1), we showed that the spot 3 (Fig. 1D ) (also indicated by an arrow in Fig. 1A ) con-tains both fragments: the spot hybridized to the probes corresponding to either of them (Fig. 1A and B ). However, the com-plex appears at the position that migrates slower than unreplicated plasmid in the second dimension upon AflI I I and ScaI digest when both repeats belong to the same fragment (Fig. 2). We suggest that this fragment contains a loop generated by the interaction between the two GAA repeats (Fig. 2C ) that slows it down in the second dimension, and has little effect on the mobility in the first dimension sinceThe method of two-dimensional elec-trophoresis 22,23 allowed us to analyze the replication progression through a DNA fragment containing two GAA repeats. In this method, the replication intermediates are isolated under non-denaturing condi-tions, digested by a restriction enzyme, and separated on two consecutive gel runs in perpendicular directions. The first direction runs in 0.4% agarose (that separates mostly by mass), and the second direction runs in 1% agarose (that sepa-rates by both mass and shape of a DNA molecule).two GAA stretches. Two GAA repeats has been shown to readily form complexes, such as “sticky DNA,” in vitro,16 but it is not obvious whether it can also form inside the mammalian cells. The sticky DNA requires more than one GAA stretch to form.21 We studied the interaction of two GAA repeats located within the same plas-mid, but since the human genome contains at least several hundreds of long GAA motif repeats,12,13 this structure can theoretically form in the genomic environment as well. However, more experiments are needed to detect its formation in genome.Figure 1. A complex between two (GAA)57 repeats within the same plasmid. Plasmid replication intermediates were isolated from Cos-1 cells 30 h after transient transfection. Intermediates were digested by restriction enzymes indicated in the plasmid maps, and separated by two-dimension-al neutral-neutral agarose gel electrophoresis as described previously.1 The gel was transferred to a nylon membrane and hybridized to one of the probes indicated in the plasmid maps as green lines. A position of the complex at the 2D gel pattern depends on the restriction digest of the replication intermediates (indicated by a red arrow). (A ) Two-dimensional electrophoresis of replication intermediates digested by PvuII that places each repeat within a separate fragment. The membrane was hybridized with probe 1 indicated in (C ). The names of the plasmids GAAGAA, GAACTT, etc., reflect the orientation of the GAA repeats within the plasmid (GAAGAA means that the two GAA stretches are in the direct orientation). (B ) The same membrane was stripped of probe 1, and re-hybridized with probe 2 (C ). (C ) The scheme of the plasmid that was used in the experiment in (A and B ). Two different fragments that resulted from the PvuII digest are shown in red and blue. The positions of GAA repeats are shown in black. They can be in the direct or the reverse orientations in this plasmid. (D ) The scheme of the 2D gel in (A ). Spot 1: unreplicated blue fragment, spot 2: unreplicated red fragment that appears because of the cross-contamination of probe 1 with probe 2 due to their preparation from the same plasmid with a restriction digest. Spot 3: a complex between the red and the blue fragments. Spikes 3-2 and 3-4 may result from double-stranded breaks in the plasmid during transfection that make one of the arms of the complex in spot 3 shorter.it has the same mass as the unreplicated fragment.The complexes formed only when the two GAA stretches were positioned on a plasmid as direct repeats (GAAGAA and CTTCTT in the plasmid names). The inverted repeats GAACTT and CTTGAA did not form complexes as shown in Figures 1 and 2. This is in agreement with the sticky DNA formation in supercoiled plasmids containing two GAA repeats that has been previously shown in vitro.16 In sticky DNA, the two GAA strands that are in the antiparallel orientation, and the CTT strand, form a stable complex stabilized by Mg2+-dependent reverse-Hoogsteen triads. However, the sticky DNA complex fell apart upon heating in the presence of EDTA, which removes the Mg2+ ions necessary for its stability,16 while we did not detect any changes in spot 3 upon heating the intermediates with EDTA (Fig. 3B). We suggest that in our case the complex may be different from the canonical sticky DNA. It may be based on Hoogsteen base pairing where the Mg2+ is not needed and a slightly acidic pH has a stabilizing effect.24 It has been shown that this type of structure forms within long GAA stretches in vitro even at a pH that is close to neutral.15 We also cannot exclude that the complexes are hemicatenated molecules connected at GAA repeats with Watson-Crick pairing.25The complexes between the two GAA repeats persisted only until the plasmids went through one replication round. DpnI restriction enzyme is a frequent-cutter that digests all DNA that contains strands syn-thesized in bacteria: it cleaves DNA that is methylated at GATC by dam methyl-ase, which is only present in bacteria, but not in mammalian cells. Extensive DpnI digest that we performed, cleaved the ini-tial DNA used in transfection, as well as the products of the first replication cycleA complex between the two repeats within the same fragment slow down its progres-sion in the second dimension of the 2D gel. The same plasmid as in the Figure 1 was used in this experiment, however, they were digested with different enzymes. (A) Replication intermediates were digested with AflIII and ScaI, placing both repeats within the same fragment. The complex of two GAA repeats results in a slowly migrating structure that is shown by a red arrow. (B) A map of the digest of the same plasmid as in Figure 1 with restriction enzymes ScaI and AflIII. Here both of the repeats are located within the same fragment shown in blue. (C) The scheme of the 2D gel in . Spots 1 and 2 are the same as in Figure 1D. Spot 3: a looped intermediate that resulted from the interaction of the two GAA repeats.A complex between the two GAA repeats does not form in plasmids that went through more than two replication rounds in mammaliancells. Two-dimensional gels of replication intermediates of a plasmid containing two (GAA)57 repeats were obtained as described in the Figure 1) Two-dimensional gel of replication intermediates digested by AflIII (placing the two repeats at two different fragments). A red arrow indi-cates the position of the complex between the two GAA-containing fragments. (B) The same replication intermediates were incubated at 80°C in the presence of 10 mM EDTA for 10 min; the pattern of the 2D gel did not change. (C) The same intermediates were additionally digested with10 units of DpnI for 2 h prior to loading. The spot at the position indicated by an arrow in Figure 1 A is not present in this picture. An additional spot that appeared in this pattern is likely not a part of the pattern, and is probably due to some contamination. (D) A map of the plasmid that was used inFigure 1A and B. Spot 1, unreplicated blue fragment; spot 2, unreplicated red fragment (which appeared due to contamination of probe 1 with other plasmid sequences). Spot 3, a complex between the blue and the red fragments. A very faint duplicate Y arc from replication of the second fragment originates from spot 2. The spikes originating from spot 3 can be interpreted the same way as11. Ku S, Soragni E, Campau E, Thomas EA, Altun G, Laurent LC, et al. Friedreich’s ataxia induced plu-ripotent stem cells model intergenerational GAA·TTC triplet repeat instability. Cell Stem Cell 2010; 7:631-7; PM I D:21040903; /10.1016/j.stem.2010.09.014.12. Siedlaczck I , Epplen C, Riess O, Epplen JT. Simple repetitive (GAA)n loci in the human genome. Electrophoresis 1993; 14:973-7; PM I D:7907288; /10.1002/elps.11501401155.13. Chauhan C, Dash D, Grover D, Rajamani J, MukerjiM. Origin and instability of GAA repeats: insights fromAlu elements. J Biomol Struct Dyn 2002; 20:253-63;PMID:12354077; /10.1080/07391102.2002.10506841.14. Gacy AM, Goellner GM, Spiro C, Chen X, Gupta G, Bradbury EM, et al. GAA instability in Friedreich’sAtaxia shares a common, DNA-directed and intraal-lelic mechanism with other trinucleotide diseases. MolCell 1998; 1:583-93; PMI D:9660942; http://dx.doi.org/10.1016/S1097-2765(00)80058-1.15. Potaman VN, Oussatcheva EA, Lyubchenko YL, Shlyakhtenko LS, Bidichandani SI, Ashizawa T , et al.Length-dependent structure formation in Friedreich ataxia (GAA)n*(TTC)n repeats at neutral pH. NucleicAcids Res 2004; 32:1224-31; PMID:14978261; http:///10.1093/nar/gkh274.16. Sakamoto N, Chastain PD, Parniewski P , OhshimaK, Pandolfo M, Griffith JD, et al. Sticky DNA: self-association properties of long GAA.TTC repeats inR.R.Y triplex structures from Friedreich’s ataxia. MolCell 1999; 3:465-75; PMID:10230399; http://dx.doi.org/10.1016/S1097-2765(00)80474-8.17. Heidenfelder BL, Makhov AM, Topal MD. Hairpinformation in Friedreich’s ataxia triplet repeat expansion.J Biol Chem 2003; 278:2425-31; PMI D:12441336;/10.1074/jbc.M210643200.18. LeProust EM, Pearson CE, Sinden RR, Gao X. Unexpected formation of parallel duplex in GAA and TTC trinucleotide repeats of Friedreich’s ataxia. J MolBiol 2000; 302:1063-80; PM D:11183775; http:///10.1006/jmbi.2000.4073.19. McIvor EI, Polak U, Napierala M. New insights into repeat instability: role of RNA•DNA hybrids. RNABiol 2010; 7:551-8; PMI D:20729633; http://dx.doi.org/10.4161/rna.7.5.12745.20. Chandok GS, Kapoor KK, Brick RM, Sidorova JM, Krasilnikova MM. A distinct first replication cycleof DNA introduced in mammalian cells. NucleicAcids Res 2011; 39:2103-15; PMID:21062817; http:///10.1093/nar/gkq903.21. Sakamoto N, Ohshima K, Montermini L, Pandolfo M, Wells RD. Sticky DNA, a self-associated complexformed at long GAA*TTC repeats in intron 1 of thefrataxin gene, inhibits transcription. J Biol Chem2001; 276:27171-7; PMI D:11340071; http://dx.doi.org/10.1074/jbc.M101879200.22. Krasilnikova MM, Mirkin SM. Analysis of tripletrepeat replication by two-dimensional gel electro-phoresis. Methods Mol Biol 2004; 277:19-28;PMID:15201446.23. Friedman KL, Brewer BJ. Analysis of replicationintermediates by two-dimensional agarose gel elec-trophoresis. Methods Enzymol 1995; 262:613-27; PM I D:8594382; /10.1016/0076-6879(95)62048-6.24. Frank-Kamenetskii MD, Mirkin SM. T riplex DNA structures. Annu Rev Biochem 1995; 64:65-95; PM D:7574496; /10.1146/annurev.bi.64.070195.000433.25. Lucas I, Hyrien O. Hemicatenanes form upon inhibition of DNA replication. Nucleic Acids Res 2000; 28:2187-93; PM D:10773090; /10.1093/nar/28.10.2187.26. McLay DW, Clarke HJ. Remodelling the paternal chromatin at fertilization in mammals. Reproduction 2003; 125:625-33; PM D:12713425; /10.1530/rep.0.1250625.because they contain one strand synthe-sized in bacteria.The replication intermediates digested with DpnI did not contain the spot 3, cor-responding to the complex between two GAA stretches (Fig. 3C ). We suggest that the absence of the complex is due to the chromatin coverage of the plasmid that accompanies replication. This is similar to our observation that GAA repeats only block replication during the first replica-tion round, until the chromatin is formed. The replication blockage that we have pre-viously observed is consistent with forma-tion of a triplex that occurs in transfected DNA only prior to nucleosome cover-age.1 Here, the complex between the two (GAA)57 repeats also occurred only with-out the chromatin structure. The absence of the complex in replicated DNA also shows that the complexes that we observe are not an artifact of the isolation and sub-sequent treatment of our intermediates, since then they would exist in at least some fraction of the replicated DNA as well.The question remains whether the non-B DNA structures can form within GAA repeats in mammalian cells since their formation requires DNA stretches that are not folded in chromatin. A win-dow when these complexes can form during development is the spermatogen-esis when the maturing sperm chromatin changes from nucleosome- to protamine-bound assembly.26 Another opportunity to form complexes comes when the chroma-tin of a sperm and an egg restructure after the fusion of the gamets.26,27 This is asso-ciated with degradation of protamines and nucleosome deposition, as the zygote DNA may lack a compact chromatin structure.28 It should be noted that the expansions in Friedreich ataxia were traced to the early divisions of the zygote.7An opportunity for the complexes to form may also exist in cancer cells. It is known that some regions of their genome are overmethylated and convert in hetero-chromatin, while other regions are under-methylated, which may promote a loose chromatin packaging.29,30It is not clear whether the two-repeats complexes would compete with triplex structure formation within each individ-ual (GAA)57 repeat. It is possible that both of them exist and contribute to overallgenomic instability. However, a separate study is necessary to determine whether these structures indeed have a biological role.Disclosure of Potential Conflicts of InterestNo potential conflicts of interest weredisclosed.Acknowledgments This study was supported by NI H grant GM087472, and research grant from Friedreich’s Ataxia Research Alliance toMMK.References 1. Chandok GS, Patel MP , Mirkin SM, Krasilnikova MM.Effects of Friedreich’s ataxia GAA repeats on DNAreplication in mammalian cells. Nucleic Acids Res2012; 40:3964-74; PM D:22262734; http://dx.doi.org/10.1093/nar/gks021.2. Kelkar YD, Tyekucheva S, Chiaromonte F , MakovaKD. The genome-wide determinants of humanand chimpanzee microsatellite evolution. GenomeRes 2008; 18:30-8; PMI D:18032720; http://dx.doi.org/10.1101/gr.7113408.3. Sharma R, Bhatti S, Gomez M, Clark RM, MurrayC, Ashizawa T, et al. The GAA triplet-repeat sequencein Friedreich ataxia shows a high level of somaticinstability in vivo, with a significant predilection forlarge contractions. Hum Mol Genet 2002; 11:2175-87; PM I D:12189170; /10.1093/hmg/11.18.2175.4. Pandolfo M. The molecular basis of Friedreichataxia. Adv Exp Med Biol 2002; 516:99-118;PM D:12611437; /10.1007/978-1-4615-0117-6_5.5. Pandolfo M. Friedreich ataxia. Arch Neurol 2008;65:1296-303; PM I D:18852343; http://dx.doi.org/10.1001/archneur.65.10.1296.6. Campuzano V, Montermini L, Moltò MD, PianeseL, Cossée M, Cavalcanti F , et al. Friedreich’s ataxia:autosomal recessive disease caused by an intronic GAAtriplet repeat expansion. Science 1996; 271:1423-7; PM ID:8596916; /10.1126/sci-ence.271.5254.1423.7. De Michele G, Cavalcanti F , Criscuolo C, Pianese L,Monticelli A, Filla A, et al. Parental gender, age at birthand expansion length influence GAA repeat intergener-ational instability in the X25 gene: pedigree studies andanalysis of sperm from patients with Friedreich’s ataxia.Hum Mol Genet 1998; 7:1901-6; PMI D:9811933; /10.1093/hmg/7.12.1901.8. De Biase I , Rasmussen A, Monticelli A, Al-MahdawiS, Pook M, Cocozza S, et al. Somatic instability of theexpanded GAA triplet-repeat sequence in Friedreich ataxia progresses throughout life. Genomics 2007;90:1-5; PMID:17498922; /10.1016/j.ygeno.2007.04.001.9. De Biase I , Rasmussen A, Endres D, Al-Mahdawi S,Monticelli A, Cocozza S, et al. Progressive GAA expan-sions in dorsal root ganglia of Friedreich’s ataxia patients.Ann Neurol 2007; 61:55-60; PMID:17262846; http:///10.1002/ana.21052.10. Du J, Campau E, Soragni E, Ku S, Puckett JW,Dervan PB, et al. Role of mismatch repair enzymesin GAA·TTC triplet-repeat expansion in Friedreichataxia induced pluripotent stem cells. J Biol Chem2012; 287:29861-72; PMID:22798143; http://dx.doi.org/10.1074/jbc.M112.391961.29. Watanabe Y, Maekawa M. Methylation of DNAin cancer. Adv Clin Chem 2010; 52:145-67; PM D:21275343; /10.1016/S0065-2423(10)52006-7.30. Kulis M, Esteller M. DNA methylation and cancer.Adv Genet 2010; 70:27-56; PMID:20920744; /10.1016/B978-0-12-380866-0.60002-2.27. Spinaci M, Seren E, Mattioli M. Maternal chromatinremodeling during maturation and after fertilization in mouse oocytes. Mol Reprod Dev 2004; 69:215-21; PM ID:15293223; /10.1002/mrd.20117.28. I mschenetzky M, Puchi M, Gutierrez S, MontecinoM. Sea urchin zygote chromatin exhibit an unfolded nucleosomal array during the first S phase. J Cell Biochem 1995; 59:161-7; PM D:8904310; /10.1002/jcb.240590205.。

weaver的分子生物学__解释说明

weaver的分子生物学解释说明1. 引言1.1 概述分子生物学是研究生命体内分子结构、功能和相互关系的科学领域。

它涉及到DNA、RNA、蛋白质等生物大分子的结构、功能以及它们在细胞中发挥的作用。

近年来，随着生物技术的快速发展，分子生物学已经成为许多领域中不可或缺的一部分，如基因工程、蛋白质工程和药物研发等。

本文将重点介绍Weaver对分子生物学的重要贡献和影响。

Weaver是一位杰出的科学家，他在分子生物学领域做出了巨大的贡献，并且推动了该领域的发展。

1.2 文章结构本文将按照以下顺序展开内容：首先介绍Weaver的分子生物学历史背景，包括他个人背景以及对这一领域做出的重要贡献。

然后，我们将详细解释Weaver 用于研究对象与方法，并探讨其在分子生物学方面取得的进展和成果。

接下来，我们将介绍在分子生物学中的关键概念，包括DNA结构与功能、基因表达和调控机制，以及蛋白质合成和折叠过程。

然后，我们将详细阐述Weaver在分子生物学中的贡献，包括发现重要基因及其功能解析、揭示DNA复制与修复机制以及探索基因调控网络和蛋白质相互作用网络等方面。

最后，我们将总结Weaver的贡献和影响力，并展望他在未来研究中的价值与应用前景。

1.3 目的本文的目的是通过对Weaver在分子生物学领域所做出的重要贡献进行全面解读和说明。

旨在向读者传达Weaver对于分子生物学发展的重要性，并强调他对该领域取得进展所做出的不可或缺的贡献。

通过深入了解Weaver的工作，我们可以更好地理解分子生物学这一科学领域以及其在解决许多医学、农业和环境问题上所起到的关键作用。

以上为文章“1. 引言”部分内容，请根据需要进行修改和调整。

2. Weaver的分子生物学2.1 Weaver的历史分子生物学家Weaver，全名约翰·T·韦弗，是20世纪下半叶最重要的分子生物学家之一。

他于1928年出生在美国，曾就读于加州理工学院并获得博士学位。

抗冻机制ICE-CBF-COR途径及抗冻相关蛋白的研究文献综述和外文翻译

Stockinger等采用酵母单杂交(yeast one-hybrid)方法从拟南芥中分离出一个cDNA，其编码产物能与CRT/DRE特异结合，且在酵母系统中可激活含有CRT/DRE作为上游激活子序列的报告基因的转录(Stockinger et al.，1997).所以，此蛋白具有转录激活因子的作用，被命名为CRT/DRE结合因子CBF1，Southern杂交结果表明，CBFI是一个单拷贝或低拷贝数基因.以包dina et al., 1999; Liu et al.，2002).与此同时，在独立进行的另一研究中，Liu等鉴定出5种DRE结合蛋白，其中DREBIA、DREBIB和DREBIC分别对应于CBF3、CBF1和
Yamaguchi Shinozaki和Shinozaki(1994)从RD29A基因的启动子中鉴定出一个9bp(TACCGACAT)的DNA调控元件DRE(dehydration-responsive element)，(Yamaguchi-Shinozak and Shinozal，1994).同年，从COR15基因的启动子中又鉴定出另一调控元件CRT(C-repeat，TGGCCGAC)(Tbata et al.，2000).这两个元件均含有CCGAC核心序列，即LTRE(low-temperature-responsive element)元件(Jiang et al.，1996).随后证明，CRT/DRE或其核心序列普遍存在于冷诱导和脱水诱导基因的启动子中，为这类基因的冷诱导和脱水诱导表达所必需.
冷调节基因（COR）的启动子区域含有具有5个碱基核心序列CCGAC的脱水反应元素DRE(Dehydration responsive element)做为顺式作用启动子元素，能在低温胁迫时激活COR基因的表达[3]。Masakazu H等将Cu2COR19转入烟草，和对照烟草植株相比较，转基因植株电渗值较低、发芽率较高、抗冻性有所提高口[4]。在这些基因表达的产物中，一些已证实是有己知酶活性的蛋白，它们可能对提高抗寒性有一定程度的作用,如拟南芥冷诱导基因中的fad8基因[5]和大麦中的blt4基因[6]，它们分别编码脂肪酸去饱和酶和一种脂肪迁移蛋白，通过这种脂肪酸去饱和酶或脂肪迁移蛋白改变质膜的组成以提高膜的冷稳定性。

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在旱地土壤中产甲烷古菌活性对养牛业的影响维维安radl1,5，安德烈亚斯gattinger1,5，艾莉卡时ˇ一´可娃´2,3，安娜NEˇmcova´2,3，Jiri Cˇuhel2,3，米洛斯拉夫的ˇimek2,3，让查尔斯munch1,4，迈克尔schloter4和Dana elhottova´21土壤生态学，慕尼黑工业大学，上施莱斯海姆，慕尼黑，德国；2生物中心，土壤生物学研究所，Cˇ艾斯克´不得ˇjovice，捷克共和国；3生物科学，南波西米亚州大学，Cˇ艾斯克´不得ˇjovice，捷克共和国；4gsf国家研究中心环境与健康，土壤生态，Neuherberg学院，德国。

在本研究中，我们测试的假设是动物的行走与作为越冬牧场土壤中的甲烷有机物有关。

因此，捷克共和国指出，在波西米亚南部的一个农场中，甲烷排放量和产甲烷菌种群对牛有不同程度的影响。

在春天，甲烷排放与动物影响的梯度相一致。

分析应用磷脂，该古细菌量最高的影响，发现部分（SI）对其有影响，其次是温和的影响（MI）没有影响。

对于产甲烷菌的实时显示甲基辅酶M还原酶（MCRA）基因的定量PCR分析观察到了相同的趋势。

检测单不饱和脂肪酸异戊烯基侧链的碳氢化合物（i20:1）表示的乙酸分解的存在影响牛产甲烷菌。

这个结果是由mcrA基因序列分析证实得到的，这表明，所分析的克隆的33%属于甲烷。

克隆序列的大部分（41%）与未培养瘤胃有关。

由此可得到的假设是，相当大的一部分来自放牧本身产生甲烷的区域。

相比于春天采样，在秋天，古细菌的生物量和mcrA数显著减少主要用于截面MI基因观察。

可以得出结论，5个月后没有牛的影响，严重影响了部分保持其产甲烷的潜力，而在温和的冲击后甲烷生产潜力。

期刊名称（2007）1，443，452–；DOI: 10.1038/ismej.2007.60；网上公布19七月2007学科类别：微生物生态学和自然栖息地的功能多样性关键词：多样性；甲烷排放；甲基辅酶M还原酶引言农业对于在土壤和植物生物量的二氧化碳（OCA，2006）气体减排和系统隔离提供了巨大的潜力。

在这方面，山地草原在低投入农业系统中被认为是作为温室气体甲烷（CH4）（胡¨tsch等人。

，1994）和只有微弱的来源的氧化亚氮（N2O）（莫西尔等人。

，1991）。

然而，当草原用作牧场放牧时这些减排潜力可以改变。

例如，克莱顿等人（1994）发现，一个放牧的牧场的N2O排放量比不放牧的草地高三倍，并且推测踩踏和排泄的相互作用会刺激这个过程。

动物踩踏时通过土壤压实造成土壤通气减少，从而增强的反硝化率（menneer等人，2005）。

此外，踩踏可以使丰富的有机碳物质从粪便进入土壤，刺激微生物代谢，从而增加在较低的土壤深度氧的需求（Sˇimek 等人，2006）。

厌氧环境和可用的有机碳可能有利于甲烷碳。

事实上，一些研究表明，在施用有机肥后，甲烷排放量的时间会增加。

虽然在畜牧业中甲烷通量变化对土壤强度有影响，在这样的环境中的古菌群落中，甲烷的结构和功能的数据可能会丢失。

因此，这一客观研究的目的是确定畜牧业中对甲烷排放量对土壤的丰富性和多样性在产甲烷社区室外越冬的影响。

特别是，对以下假设进行了测试：（1）牛越冬放牧导致土壤功能的变化，可导致甲烷的排放而不是甲烷的下沉，（2）排放量与动物的影响有关及（3）牛的粪便作为瘤胃产甲烷菌接种和刺激土传病原菌的生长和活动。

对于产甲烷古菌生物量的特征，我们使用了磷脂（PLEL）方法（gattinger等人，2002，2003年）。

该方法的分类分辨率相对较低，但其坚固性对不同土壤基质不一样，这已被证明（白等人，2000；瓦辛格尔。

等人，2000；瓦格纳等人，2005）。

此外，实时定量PCR和克隆库的分析甲基辅酶M还原酶（MCRA）基因，这对甲基辅酶M还原酶的A亚基编码，进行了更详细的调查研究对于产甲烷的结构。

这种酶催化还原甲基辅酶M导致释放甲烷（埃等人，1988）。

mcrA 基因的存在限制产甲烷古菌（thauer，1998）；因此，它的数量作为一个土壤中的产甲烷生物量估算。

在本研究中，我们表明，在越冬时使用的急剧变化的草原土壤的性质，这反过来又影响土壤产甲烷菌的数量和活性。

材料和方法试验场在南波西米亚捷克共和国（纬度481520 N，东经141130 E）对位于博罗瓦农场附近过冬的牛进行了调查。

约4公顷的面积已被约90头母牛从1995年（详情见ˇimek等人，2006）用于越冬（十月/十一月至四月/五月）。

土壤是沙质壤土，分类为雏形土，含有60–80%砂，14–32%淤泥和6–14%粘土（根据美国农业部的分类系统）。

植被是一种多年生的混合物草，三叶草和其他双子叶植物。

长期平均年降水量是650mm和年平均气温71C（从位于气象站7公里实验农场测得的数据）。

在冬季结束的时候（4月至5月），有一个由于对畜牧业的可见梯度的影响。

对越冬场所的影响程度与整个牧场不相同。

我们研究了这种梯度的三个部分：最受影响的地区位于牲口棚的附近（有1/4严重影响），影响较小的区域是梯度的中间部分（有1/4温和的影响）和不受影响的地区在越冬区的相对侧（部分1/4没有影响）。

在春天，研究区域的特点如下：硅完全被植被破坏，通过粪便破坏了土壤结构和饱和度；温和的影响是通过部分植被破坏和保留了土壤表面的粪便；影响小的是通过覆草和保留了不可见的粪便。

甲烷通量的测量和土壤取样该实验进行了两次，在春季（2005年5月11日），从动物从越冬区域移在秋季（2005年10月18日），从它们回来之前。

在每个区域的不同部分随机选择了一些区域进行了气体流量测量和土壤取样独立的区域可以重复在这项研究中。

甲烷通量决定在中间部分使用（断面积0.076m2，体积15dm3）。

在测量之前立刻推进到土壤3厘米处。

每个区域有一个橡胶隔膜立即插入之后进行安装。

顶部的气体样品在60分钟后被收集在室内。

初步调查显示，气体浓度在封闭区域是线性增加的。

在每个部分采集九种气体样品，存储在预先抽真空的3.5ml的小玻璃瓶中，立即送到实验室进行分析。

使用惠普5890系列II气相色谱仪（休利特帕卡德，帕洛阿尔托，CA，USA）装备用2M Porapak N列在7501c，和火焰电离检测器进行甲烷数量的测定。

仪器的校准用CH4的标准混合物，对于甲烷气体从玻璃瓶转移时的损失其结果正确的。

与气体采样相比较，真实的土壤样品是从四个区域获得的（0–20厘米深），通过采集七个附属样品，从直径为3厘米的土壤区域随机采取。

土壤立即通过5毫米的网格进行均匀化筛分。

对样品进行核酸和磷脂的分析并立即分别存储在 801c和41。

由于在分析的这些区域观测到很不均匀，因此重复测量了大多数气体。

土壤分析在1051度进行干燥，土壤的PH值使用在1:2.5（w/w）土壤/水悬浮液中的玻璃电极测量。

土壤中的无机氮（NH4þ，NO3）通过从1M氯化钾提取使用的土壤（新鲜现场潮湿的40克溶液比：200毫升（ZBı´RAL等人，1997）测量。

总有机碳测定采用重铬酸湿式氧化法（杰克逊，1958），采用凯氏定氮法测定总氮含量（ZBı´RAL，1995）。

总生物量和古细菌的测定采用PLFA法和PLEL法从新鲜土样中提取的脂类相当于10克干重（干重），根据布莱–代尔方法描述（zelles和白，1993）。

由此产生的脂质材料分离成中性脂质，糖脂和在二氧化硅的磷脂键合（spe-si；Bond Elute，分析化学国际，美国）分别通过洗脱氯仿和丙酮甲醇。

磷脂的等分试样分数相当于2.5克干重为磷脂脂肪酸（PLFA）分析。

在温和的碱性水解后，oravecz等人进行了详细描述（2004年），所得到的脂肪酸甲酯利用自动识别系统在毛细管气相色谱分离（Agilent 6850，氢火焰离子化检测器，tsba50，MIDI公司，纽瓦克，德，美国）。

另一部分的磷脂馏分相当于7.5克土壤干重采用PIEL法分析根据gattinger 等人的分析（2003年）。

在乙醚核心脂质形成后，醚连接类异戊二烯释放后裂解醚键与HI和还原脱卤在冰醋酸锌。

由此产生的类异戊二烯烃溶解在100毫升标准溶液内（十九烷甲基酯）进行气相色谱-质谱联用在操作条件进行了光谱分析（gattinger等人，2003）。

PLFA / PLEL个别化合物数据（用1 nmol克土壤干重表示）被添加到获得的总磷脂浓度链，土壤中微生物总量（zelles，1999）。

PLFA/PLEL化合物总浓度磷脂链的百分比表示，给出了一定微生物组的相对丰度估计（zelles，1999）。

以下简称为不同PLEL派生的类异戊二烯烃：i20:0表明饱和和单不饱和i20:120个碳原子的类异戊二烯链i40:0；表示一个类异戊二烯链的40个碳原子。

从土壤中提取DNA从500mg土壤中提取DNA被格利菲斯等人描述（2000年）。

提取物的数量和质量采用分光光度法测定（纳米微滴，peqlab，埃朗根，德国）。

实时定量PCR的mcrA基因实时定量PCR（qPCR）在ABI PRISM 7700序列检测系统进行（珀金埃尔默，福斯特市，CA，USA）。

反应混合物含有5毫升的qPCR ROX 和Go Green （qbiogene，伊尔基希，法国），1.5毫克牛血清蛋白（西格马-奥德里奇，德国），每种物质5mol（卢顿等人，2002），5%二甲基亚砜（西格马-奥德里奇，斯德海姆，德国），0.5毫升的DNA模板和水的最终体积25毫升。

由初始951度变性15 min和941度变性1min4放大到在721度和521度各 1分钟。

标准曲线是通过使用10倍的稀释部分稀释马氏甲烷八叠球菌mcrA基因序列建立的。

计算出的PCR扩增效率使用的数据来自标准曲线和下面的公式：[10( 1/slope)] 1。

DNA的提取物质影响废弃物质，对每个样品进行稀释。

每个样品进行四个独立的测定。

通过放大熔融的PCR产物的质量曲线和确认用1.5%溴化乙锭染色的琼脂糖凝胶。

mcrA基因的克隆和测序从第四提取物中提取五十微克的复制为模板对于mcrA基因的扩增。

对以上所描述的反应进行了混合和放大，除了聚合酶以外（Taq聚合酶，Invitrogen公司，卡尔斯鲁厄，德国）。

纯化的PCR产品从四个重复汇集和克隆TA克隆试剂盒（Invitrogen公司），按照制造商的说明。

反应中PCR产物中Ca 50纳克，缓冲液1毫升，2.5毫升的无菌蒸馏水，2毫升矢量（PCR 2.1）和T4连接酶1毫升。

白色的殖民地，包括插入，接种50 MLML 1卡那霉素Luria–金氏培养基，一夜之间长大和用于质粒分离（Qiagen质粒抽提试剂盒，QIAGEN，希尔登，德国）。

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