Cloning, characterization and expression in Escherichia coli of a leucine biosynthetic gene from Streptomyces rochei.
Leucine and histidine biosynthetic genes from Streptomyces rochei HP1 that complemented auxotrophic mutations in S. lividans TK54 were cloned in pIJ61. DNA from one leucine recombinant plasmid was subcloned into pBR322. From the latter, a recombinant plasmid was obtained that complemented the leuA mutation in Escherichia coli CV512 but not other leucine markers in E. coli. Analysis of this and several subclones, including mutant plasmids constructed in vitro, established that the cloned S. rochei gene was expressed in E. coli from the tetracycline promoter of pBR322 to produce a polypeptide of 67 kDa; the corresponding coding region was shown to be within a 1.7 kbp DNA fragment. Blot hybridization revealed corresponding homologous genes in several other streptomycetes.
Hercomb
J
J
Department of Biochemistry, University of Leeds, UK.
Thierbach
G
G
Baumberg
S
S
Parish
J H
JH
D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004274:DNA, Recombinant / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D006639:Histidine / Q000235:genetics; D007930:Leucine / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids; D012689:Sequence Homology, Nucleic Acid; D013302:Streptomyces / Q000235:genetics*
D004269:DNA, Bacterial; D004274:DNA, Recombinant; D006639:Histidine; D007930:Leucine