title: Cloning of Bacillus subtilis leucina A, B and C genes with Escherichia coli plasmids and expression of the leuC gene in E. coli.
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. beta-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.
mesh_terms: D015079:2-Isopropylmalate Synthase / Q000235:genetics / Q000378:metabolism; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics*; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D007930:Leucine / Q000235:genetics*; D010957:Plasmids
chemical_list: D004269:DNA, Bacterial; D004274:DNA, Recombinant; D015079:2-Isopropylmalate Synthase; D007930:Leucine