Chapter 7 Genetic Manipulation
第七章 遗传操作
? DNA manipulation DNA的遗传操作
? Basic techniques used to clone genes
? Construction and screening of genomic libraries
? The manipulation of cloned DNA sequences in vitro
? The molecular analysis of DNA
? Cell manipulation 细胞的遗传操作
? Human gene therapy
? Transgenic animals
DNA manipulation DNA的遗传操作
Recombinant DNA techniques
目的基因 (the gene of interest)的获取
目的基因与载体结合成重组 DNA
重组 DNA分子引入受体细胞并表达
转化体细胞的扩增、鉴定与筛选
Basic techniques used to clone genes
? The discovery of restriction endonucleases
限制性核酸内切酶
? The production of recombinant DNA
molecules in vitro
? Amplification of recombinant DNA
molecules in cloning vectors
Restriction endonucleases 限制性核酸内切酶
? 限制与修饰作用
? 限制性核酸内切酶 restriction endonucleases
E.coli K
E.coli B
E.O.P = 1E.O.P = 10-4
E.O.P = 1
λ,B
λ,K
限制酶 — 能识别和切断外来的
DNA而使它失去复制能力
修饰酶 — 能使限制酶的识别序
列中的若干个碱基甲基化，限制酶
不能识别它
Features of Restriction endonucleases
o 限制性核酸内切酶可分 I型,II型和 III型三类
o 识别序列有 3个特点：由 4~6个核苷酸组成，
回文结构，可被有规律替代
o II型限制酶的切割类型,错切和平切两类
Amplification of recombinant DNA molecules
in cloning vectors
The essential features
of a cloning vector
Strategy employed in
using phage ? as a
cloning vector
Structure of a polycloning site in a cloning vector
Structure
of pBR322
Construction and
screening of
genomic libraries
The manipulation of cloned DNA sequences in vitro
Joining DNAs with
linker and adapter
molecules
The molecular analysis of DNA
— DNA sequencing
? Sanger的加减法原理：极其复杂
的核苷酸排列顺序可由简单的不
同长度的核苷酸推算出来
? 末端终止法的原理
Cell manipulation 细胞的遗传操作
----Human gene therapy
----Transgenic animals
Human gene therapy
Transgenic animals
第七章 遗传操作
? DNA manipulation DNA的遗传操作
? Basic techniques used to clone genes
? Construction and screening of genomic libraries
? The manipulation of cloned DNA sequences in vitro
? The molecular analysis of DNA
? Cell manipulation 细胞的遗传操作
? Human gene therapy
? Transgenic animals
DNA manipulation DNA的遗传操作
Recombinant DNA techniques
目的基因 (the gene of interest)的获取
目的基因与载体结合成重组 DNA
重组 DNA分子引入受体细胞并表达
转化体细胞的扩增、鉴定与筛选
Basic techniques used to clone genes
? The discovery of restriction endonucleases
限制性核酸内切酶
? The production of recombinant DNA
molecules in vitro
? Amplification of recombinant DNA
molecules in cloning vectors
Restriction endonucleases 限制性核酸内切酶
? 限制与修饰作用
? 限制性核酸内切酶 restriction endonucleases
E.coli K
E.coli B
E.O.P = 1E.O.P = 10-4
E.O.P = 1
λ,B
λ,K
限制酶 — 能识别和切断外来的
DNA而使它失去复制能力
修饰酶 — 能使限制酶的识别序
列中的若干个碱基甲基化，限制酶
不能识别它
Features of Restriction endonucleases
o 限制性核酸内切酶可分 I型,II型和 III型三类
o 识别序列有 3个特点：由 4~6个核苷酸组成，
回文结构，可被有规律替代
o II型限制酶的切割类型,错切和平切两类
Amplification of recombinant DNA molecules
in cloning vectors
The essential features
of a cloning vector
Strategy employed in
using phage ? as a
cloning vector
Structure of a polycloning site in a cloning vector
Structure
of pBR322
Construction and
screening of
genomic libraries
The manipulation of cloned DNA sequences in vitro
Joining DNAs with
linker and adapter
molecules
The molecular analysis of DNA
— DNA sequencing
? Sanger的加减法原理：极其复杂
的核苷酸排列顺序可由简单的不
同长度的核苷酸推算出来
? 末端终止法的原理
Cell manipulation 细胞的遗传操作
----Human gene therapy
----Transgenic animals
Human gene therapy
Transgenic animals
