Document Detail

Replacement of Streptomyces hygroscopicus genomic segments with in vitro altered DNA sequences.
MedLine Citation:
PMID:  3162728     Owner:  NLM     Status:  MEDLINE    
We have developed a method for gene replacement in Streptomyces hygroscopicus which permits introduction of an in vitro derived mutation carried on a plasmid into the chromosome. We constructed the plasmid pMSB212 which can replicate in S. hygroscopicus and contains the step5 gene of the bialaphos biosynthetic pathway which was inactivated by a frame-shift mutation caused by filling in the cohesive ends of the EcoR I site in the structural gene. pMSB212 was introduced into a bialaphos producer strain and by protoplast regeneration of the primary thiostrepton-resistant transformants, non-producing mutants, were obtained. Biochemical and genetical analyses indicated that these mutants were specifically blocked by introduction of the frame-shift mutation in the step5 gene on the chromosome. This method will enable us to obtain isogenic mutants of known genes and to identify new genes encoded on a cloned fragment.
H Anzai; Y Kumada; O Hara; T Murakami; R Itoh; E Takano; S Imai; A Satoh; K Nagaoka
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Journal of antibiotics     Volume:  41     ISSN:  0021-8820     ISO Abbreviation:  J. Antibiot.     Publication Date:  1988 Feb 
Date Detail:
Created Date:  1988-05-20     Completed Date:  1988-05-20     Revised Date:  2006-05-01    
Medline Journal Info:
Nlm Unique ID:  0151115     Medline TA:  J Antibiot (Tokyo)     Country:  JAPAN    
Other Details:
Languages:  eng     Pagination:  226-33     Citation Subset:  IM    
Pharmaceutical Research Laboratories, Meiji Seika Kaisha, Ltd., Yokohama, Japan.
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MeSH Terms
Chromosomes, Bacterial
DNA, Bacterial*
Electrophoresis, Polyacrylamide Gel
Genetic Engineering / methods
Genetic Vectors
Organophosphorus Compounds / biosynthesis*
Streptomyces / genetics*,  metabolism
Transformation, Bacterial*
Reg. No./Substance:
0/DNA, Bacterial; 0/Organophosphorus Compounds; 35597-43-4/bialaphos

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