Document Detail

A general method for the construction of Escherichia coli mutants by homologous recombination and plasmid segregation.
MedLine Citation:
PMID:  3112516     Owner:  NLM     Status:  MEDLINE    
A technique is presented by which mutations can be introduced into the Escherichia coli chromosome by gene replacement between the chromosome and a plasmid carrying the mutant gene. The segregational instability of plasmids in E. coli is used with high efficiency to isolate E. coli mutants. The method should be applicable to construction of mutants for any E. coli chromosomal gene provided it is dispensable, and for any E. coli strain provided it is capable of homologous recombination. The use of the method was demonstrated by constructing E. coli mutants for the glycogen branching enzyme gene (glgB) and the beta-galactosidase gene (lacZ). The results show that recombination occurs via a reciprocal mechanism indicating that the method should, in a slightly modified form, also be useful in transferring chromosomal mutations onto multicopy plasmids in vivo.
J A Kiel; J P Vossen; G Venema
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Molecular & general genetics : MGG     Volume:  207     ISSN:  0026-8925     ISO Abbreviation:  Mol. Gen. Genet.     Publication Date:  1987 May 
Date Detail:
Created Date:  1987-09-22     Completed Date:  1987-09-22     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0125036     Medline TA:  Mol Gen Genet     Country:  GERMANY, WEST    
Other Details:
Languages:  eng     Pagination:  294-301     Citation Subset:  IM    
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MeSH Terms
1,4-alpha-Glucan Branching Enzyme / genetics
Escherichia coli / genetics*
Genetic Engineering / methods
beta-Galactosidase / genetics
Reg. No./Substance:
EC,4-alpha-Glucan Branching Enzyme; EC

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