chap three cloning vector.pptVIP

Chapter 3 Gene Cloning Vector Definition of cloning vector 通过不同途径能将承载的外源DNA片段（基因）带入受体细胞，并在其中得以维持的DNA分子。 Common properties Have multiple cloning sites for DNA insertion multiple cloning sites : it has many recognition sequences of RE and the exogenous DNA fragments can be inserted in. Can bring exogenous DNA into receptor cell, and can replicate in the host cell. Carry marker genes conferring the resistance to antibiotics(抗生素) Save for receptor cell, can not be transferred into other cells except for the receptor cell. Section one Plasmid 3 replication of plasmid Plasmid DNA can replicate before every cell division. During cell division, at least one copy of the plasmid DNA is segregated to each daughter cell, assuring continued propagation of the plasmid through successive generations of the host cell. It can keep stable copies in host. 拷贝数多的称为松驰型质粒（小质粒）， 少的称为严紧型质粒（大质粒）。 4 质粒的不亲合性 能寓于同一细胞中的质粒为亲合性质粒 不能寓于同一细胞中的质粒为不亲合性质粒。 受体细胞最好无内源质粒. 5 质粒的迁移性 接合质粒：contain tra gene, can joint the host cell and receptor cell and then transfer into the receptor cell. This kind of Plasmid is very large, with few copies in host cell, it has a wide range of hosts and is not very safe. 非接合质粒： small and safe, has not the tra gene and can not transfer into other cells itself. has more copies , Usually be used as cloning vectors. 6 显性质粒与隐蔽质粒 隐蔽质粒：the properties of the receptor cells have no change when the plasmid was introduced into the cell. 显性质粒: After plasmid is transferred into receptor cell, it can confer some new properties to the receptor cells. (1) 致育性： For example: the Ti plasmid can transfer some genes into plant cells and make the transgenic plant cell grow and produce a tumour. 二、strategy for the construction of the plasmid vector with natural plasmid The nature plasmid can replicate and have more copies in receptor cells. The nature plasmid have polylinker region. polylinker region: multiple cloning site, 3、The nature plasmid have marker genes.