Molecular cloning of a D-cycloserine resistance gene from D-cycloserine-producing Streptomyces garyphalus.
A 3.5-kb DNA fragment that confers resistance to D-cycloserine (DCS) was cloned from the chromosomal DNA of a DCS-producing Streptomyces garyphalus into Streptomyces lividans by a shot-gun cloning technique. Nucleotide sequence analysis revealed the existence of four open reading frames (ORFs B, C, D, and E), together with two incomplete ORFs, A and F. By introduction of the cloned fragment into Escherichia coli, the host obtained resistance to DCS. We showed that ORF B, which consists of 903 bp, is a DCS resistance gene. The hydropathy plot analysis of a protein deduced from ORF B revealed that the protein carries membrane-integral domains spanning the membrane 10 times, which suggests that the DCS-resistance determinant may be a factor associated with DCS transport.
Matsuo
Hiroaki
H
Department of Molecular Microbiology and Biotechnology, Graduate School of Biomedical Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8551, Japan.
Kumagai
Takanori
T
Mori
Katsuhiko
K
Sugiyama
Masanori
M
D000904:Antibiotics, Antitubercular / Q000096:biosynthesis / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics; D003001:Cloning, Molecular; D003523:Cycloserine / Q000096:biosynthesis / Q000494:pharmacology*; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism
D000904:Antibiotics, Antitubercular; D001426:Bacterial Proteins; D003523:Cycloserine