园艺植物生物技术第七章优秀课件
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园艺植物生物技术第七章
§What is gene? Gene can be defined as a segment of genome with a specific sequence of nucleotides, which has a specific biological function.
TypeⅡendonuclease
❖Recognize：4~6个核苷酸，回文序列
❖Cuting result：
• a sticky ends(粘性末端)：
Pst1
EcoR1
5’-CTGCA G -3’
5’-G AATTC-3’
3’-G ACGTC -5’
3’-CTTAA G-5’
• a blunt ends(平末端)：NruⅠ
BclⅠ
G GATC C
T GATC A
C CTAG G
A CTAG T
7.1.2 DNA ligase
（DNA连接酶）
• T4 DNA ligase： join blunt ends or sticky ends • Escherichia coli DNA ligase： join sticky ends
7.1.1 Restriction Endonuclease
一类能识别双链DNA分子中某一特定核苷酸序 列，并能对核酸内部的磷酸二酯键进行切割的一 种内切核酸酶。
TypeⅠendonuclease：识别专一位点，切割不专一； TypeⅡendonuclease：识别专一位点，切割专一； TypeⅢ endonuclease：识别专一位点，切割不专一。
modification, is the direct manipulation of an organism's genome using biotechnology.
New DNA may be inserted in the host genome by first isolating and copying the genetic material of interest using molecular cloning methods to generate a DNA sequence, or by synthesizing the DNA, and then inserting this construct into the host organism.
7.1 Instrumental Enzyme of
Gene Engineering
• Restriction endonucleases (限制性内切酶) • Ligase（连接酶） • Polymerase（聚合酶） • Nuclease（核酸酶） • Terminal enzyme（末端转移酶） • Methylase（甲基化酶）
பைடு நூலகம்
5’ TCG CGA 3’
5’ AGC GCT 3’
• Isoschizomers(同裂酶)：指来源不同，但识别相同 靶序列的核酸内切酶。
如：HpaⅡ和MspⅠ是一对同裂酶 (CCGG) • Isocaudarner(同尾酶)：指来源不同、识别靶序列
不同但产生相同的粘性末端的核酸内切酶。
BamHⅠ
• Genetically modified crops (GMCs, GM crops).The first genetically modified plant was produced in 1982, using an antibiotic-resistant tobacco plant. The first genetically modified crop approved for sale in the U.S., in 1994, was the FlavrSavr tomato, which had a longer shelf life.
gene
intron intron intron
Enhancer element
Upstream element
TATA box
Exon Exon Exon
controlling sequences Structural gene
• Genetic Engineering also called genetic
7.1.3 DNA polymerase
1. DNA polymeraseⅠof E.coli：
5’→3’聚合活性， 5’→3’外切活性, 3’→5’ 外切活性
2.Klenow fragment of DNA polymeraseⅠof E.coli
5’→3’聚合活性， 3’→5’ 外切活性
3.DNA polymerase of Phage T4
Genes may be removed, or "knocked out", using a nuclease. Gene targeting is a different technique that uses homologous recombination to change an endogenous gene, and can be used to delete a gene, remove exons, add a gene, or introduce point mutations.
5’→3’聚合活性， 3’→5’ 外切活性，活性高200倍。
4.Reverse transcriptase(反转录酶) 5.DNA polymerase of T7 phage ：活性高1000倍
7.1.4 Other DNA-modifying enzymes
• Terminal transferase(末端转移酶)：能以具 3’-OH的单链和双链的DNA为引物，在有底物dNTP 和Mg2+和Co2+下，把脱氧核苷酸一个接一个加到 DNA的3’端上。
• An organism that is generated through genetic engineering is considered to be a genetically modified organism (GMO). The first GMOs were bacteria in 1973.
§What is gene? Gene can be defined as a segment of genome with a specific sequence of nucleotides, which has a specific biological function.
TypeⅡendonuclease
❖Recognize：4~6个核苷酸，回文序列
❖Cuting result：
• a sticky ends(粘性末端)：
Pst1
EcoR1
5’-CTGCA G -3’
5’-G AATTC-3’
3’-G ACGTC -5’
3’-CTTAA G-5’
• a blunt ends(平末端)：NruⅠ
BclⅠ
G GATC C
T GATC A
C CTAG G
A CTAG T
7.1.2 DNA ligase
（DNA连接酶）
• T4 DNA ligase： join blunt ends or sticky ends • Escherichia coli DNA ligase： join sticky ends
7.1.1 Restriction Endonuclease
一类能识别双链DNA分子中某一特定核苷酸序 列，并能对核酸内部的磷酸二酯键进行切割的一 种内切核酸酶。
TypeⅠendonuclease：识别专一位点，切割不专一； TypeⅡendonuclease：识别专一位点，切割专一； TypeⅢ endonuclease：识别专一位点，切割不专一。
modification, is the direct manipulation of an organism's genome using biotechnology.
New DNA may be inserted in the host genome by first isolating and copying the genetic material of interest using molecular cloning methods to generate a DNA sequence, or by synthesizing the DNA, and then inserting this construct into the host organism.
7.1 Instrumental Enzyme of
Gene Engineering
• Restriction endonucleases (限制性内切酶) • Ligase（连接酶） • Polymerase（聚合酶） • Nuclease（核酸酶） • Terminal enzyme（末端转移酶） • Methylase（甲基化酶）
பைடு நூலகம்
5’ TCG CGA 3’
5’ AGC GCT 3’
• Isoschizomers(同裂酶)：指来源不同，但识别相同 靶序列的核酸内切酶。
如：HpaⅡ和MspⅠ是一对同裂酶 (CCGG) • Isocaudarner(同尾酶)：指来源不同、识别靶序列
不同但产生相同的粘性末端的核酸内切酶。
BamHⅠ
• Genetically modified crops (GMCs, GM crops).The first genetically modified plant was produced in 1982, using an antibiotic-resistant tobacco plant. The first genetically modified crop approved for sale in the U.S., in 1994, was the FlavrSavr tomato, which had a longer shelf life.
gene
intron intron intron
Enhancer element
Upstream element
TATA box
Exon Exon Exon
controlling sequences Structural gene
• Genetic Engineering also called genetic
7.1.3 DNA polymerase
1. DNA polymeraseⅠof E.coli：
5’→3’聚合活性， 5’→3’外切活性, 3’→5’ 外切活性
2.Klenow fragment of DNA polymeraseⅠof E.coli
5’→3’聚合活性， 3’→5’ 外切活性
3.DNA polymerase of Phage T4
Genes may be removed, or "knocked out", using a nuclease. Gene targeting is a different technique that uses homologous recombination to change an endogenous gene, and can be used to delete a gene, remove exons, add a gene, or introduce point mutations.
5’→3’聚合活性， 3’→5’ 外切活性，活性高200倍。
4.Reverse transcriptase(反转录酶) 5.DNA polymerase of T7 phage ：活性高1000倍
7.1.4 Other DNA-modifying enzymes
• Terminal transferase(末端转移酶)：能以具 3’-OH的单链和双链的DNA为引物，在有底物dNTP 和Mg2+和Co2+下，把脱氧核苷酸一个接一个加到 DNA的3’端上。
• An organism that is generated through genetic engineering is considered to be a genetically modified organism (GMO). The first GMOs were bacteria in 1973.