基因克隆第6章.pptVIP

Chapter 6 Cloning Vectors for E.coli In this chapter，we study： Chapter 6 Cloning Vectors for E.coli 6.1 Cloning vectors based on E.coli plasmids 6.1 Cloning vectors based on E.coli plasmids 6.1.1 The nomenclature of plasmid cloning vectors 6.1.2 The useful properties of pBR322 6.1.3 The pedigree of pBR322 6.1.4 Other typical E.coli plasmid cloning vectors 6.1.1 The nomenclature of plasmid cloning vectors 6.1.2 The useful properties of pBR322 Why has pBR322 been such a popular cloning vector ? 6.1.3 The pedigree of pBR322 6.1.4 Other typical E.coli plasmid cloning vectors pBR327 pBR327 differs from pBR322 in two important ways： pUC 8 —— a lac selection plasmid pUC8 has three important advantages: pGEM3Z ——in vitro transcription of cloned DNA 6.2 Cloning vectors based on λbacteriophage Two problems had to be solved: （1） The size limitation （2） Multiple restriction sites 6.2 Cloning vectors based on λbacteriophage 6.2.1 Segments of the λ genome can be deleted without impairing viability 6.2.2 Natural selection can be used to isolate modified λ that lack certain restriction sites 6.2.3 Insertion and replacement vectors 6.2.4 Cloning experiments with λinsertion or replacement vectors 6.2.5 Very large DNA fragments can be cloned using a cosmid 6.2.1 Segments of the λ genome can be deleted without impairing viability 6.2.2 Natural selection can be used to isolate modified λ that lack certain restriction sites How to solve the problem of multiple restriction sites for most restriction endonucleases ? 6.2.3 Insertion and replacement vectors 6.2.4 Cloning experiments with λinsertion or replacement vectors 6.2.5 Very large DNA fragments can be cloned using a cosmid Cosmid 6.3 λ and other high capacity vectors enable genomic libraries to be constructed How many clones are need for a genomic library? New cloning vectors to handle larger DNA inserts 1 using the circula