Cloning of Bacillus subtilis leucine A, B and C genes with Escherichia coli plasmids and expression of the leuC gene ... View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1978-01

AUTHORS

Kenji Nagahari, Kenji Sakaguchi

ABSTRACT

The leucine genes of Bacillus subtilis have been cloned directly from the chromosomal DNA into Escherichia coli leuB cells by selection for the Leu+ phenotype using RSF2124 as a vector plasmid. The hybrid plasmid designated RSF2124-B·leu contained a 4.2 megadalton fragment derived from B. subtilis DNA, including the leu genes. The fragment had one site susceptible to EcoRI* and another site susceptible to BamNI endonuclease. Among the three fragments produced by EcoRI* and BamNI endonucleases, the 1.2 megadalton fragment had the ability to transform B. subtilis leuA, leuB and leuC auxotrophs to leu+. However, B. subtilis ilvB and ilvC auxotrophs were not rescued even by the whole 4.2 megadalton fragment present in the hybrid plasmid. β-Isopropylmalate dehydrogenase (leuB gene product) activity found in E. coli cells containing the hybrid plasmid was about 60% of that in E. coli wild type cells, despite the high copy number (7.8) of the plasmid per chromosome observed. More... »

PAGES

263-270

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf00267197

DOI

http://dx.doi.org/10.1007/bf00267197

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1045410255

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/415224


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