Open AccessCloning and characterization of the gene encoding penicillin‐binding protein A of Streptomyces griseus 1
Author(s) -
Jiang Hao,
Kendrick Kathleen E.
Publication year - 2000
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2000.tb09403.x
Subject(s) - streptomyces griseus , biology , gene , cosmid , penicillin binding proteins , genetics , microbiology and biotechnology , streptomyces , bacteria , escherichia coli
An internal segment of the penicillin‐binding protein gene, pbpA , of Streptomyces griseus was amplified from genomic DNA using the polymerase chain reaction and used as a hybridization probe to isolate the complete gene from a cosmid library. pbpA encodes a 485 amino acid sequence that conserves three motifs of PBPs, SXXK, SXN, and KTG. The pbpA gene was located downstream of a gene homologous to the Bacillus subtilis spoVE gene. The pbpA gene was disrupted by replacing an Apa I fragment of the pbpA gene in S. griseus chromosome with an apramycin resistance gene cassette or directly inserting this apramycin resistance gene cassette at the Nco I site of pbpA penicillin‐binding domain. No obvious defects in growth, sporulation, or spore sonication resistance were observed in the constructed pbpA mutants, suggesting that PBPA is not essential for growth and sporulation under normal laboratory conditions in S. griseus .