曼氏无针乌贼肌肉cDNA文库的构建和表达序列标签分析

曼氏无针乌贼肌肉cDNA文库的构建和表达序列标签分析蒋霞敏;王佳;薛良义;曾建刚
【期刊名称】《应用海洋学学报》
【年(卷),期】2011(030)003
【摘要】A cDNA library from the muscle tissue of Sepiella maindroni was constructed using a cDNA library construction kit. The quality of the library reached 1. 2 x 106 cfu/dm3 in capacity while the rate of recombinant efficiency was as high as 96%. PCR results showed that the average length of the inserts was greater than 1 000 bp. 568 clones were selected and sequenced from the library, among which 475 that contained expressed sequence tags ( ESTs) were greater than 100 bp. Approximately 200 single-gene clusters were acquired including 41 contigs and 159 single-copy ESTs after an initial assembly, while the redundancy rate was 57.89%. Through retrievals, 53 (26.5% ) ESTs had no obvious homology with any known sequences in the BLASTx (E-value ≥ 1.00 x 10 -10) , and were identified as novel genes. 147 (73. 5% ) ESTs had relatively high homology with previously reported genes, including energy-related genes with the highest abundance of ESTs (23.0% ), cellular signal transduction ( 17.5% ) , cytoskeleton (12. 5% ) , and protein metabolism (9. 5% ) , in addition to heat shock protein 70 and transferrin. These ESTs might facilitate further screening and cloning of uniquely expressed genes in the muscle tissues of Sepiella maindroni and other cephalopods.%以曼氏无针乌贼(Sepiella
maindroni)肌肉组织为研究材料,利用Clontech公司的cDNA文库构建试剂盒构建了曼氏无针乌贼肌肉cDNA文库.对文库质量的分析结果表明:cDNA文库的库容量约为1.2×106克隆(cfu/dm3),重组率达96％,插入片段平均长度大于1 000 bp.挑取568个阳性克隆进行5’末端测序,其中475条ESTs长度大于100 bp,初步拼接后得到200个单基因簇,包括41个重叠群和159个单拷贝EST,冗余度为57.89％.经检索,53条ESTs(占比为26.5％)在BLASTx上无明显的同源性(E值
≥1.00×10 -10),为新基因.147条ESTs(占比为73.5％)与已报道的基因有较高的同源性；其中与能量相关基因的ESTs丰度最高,占总数的23.0％.细胞信号传导、细胞骨架和蛋白质代谢的次之,比例分别为17.5％、12.5％、9.5％,其中包括热激蛋白70、转铁蛋白等.这些EST数据有助于进一步筛选和克隆曼氏无针乌贼和其他头足动物的肌肉特异性表达基因.
【总页数】6页(P357-362)
【作者】蒋霞敏;王佳;薛良义;曾建刚
【作者单位】宁波大学生命科学与生物工程学院,浙江宁波315211;宁波大学生命科学与生物工程学院,浙江宁波315211;宁波大学生命科学与生物工程学院,浙江宁波315211;宁波大学生命科学与生物工程学院,浙江宁波315211
【正文语种】中文
【中图分类】Q178.53
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