具有产生安莎类抗生素潜能放线菌分子筛选_英文_.pdfVIP

396 2008 7 33 7 : 1001-8689( 2008) 07-0396-07 Molecular screening of potential ansamycin producers from actinomycetes Wu Lin-zhuan, Zhang Hui-tu, Han Feng, Gao Qun-jie, un Gui-zhiandWang Yi-guang ( , ; , Key Laboratory of Biotechnology of Antibiotics M inistry of Health Inst itute of Medicinal Biotechnology CAM PUMC, Beijing 100050) ABSTRACT3-amino-5-hydroxybenzoic acid ( AHBA ) is the specific starter unit for all ansamycin biosyn- thesis. AHBA synthase is the unique terminal enzyme converting 5-amino-3-dehydro-5-deoxyshikimate into AHBA in the aminoshikimate pathw ay specifically for AHBA biosynthesis. AHBA synthase gene could be used - - . - as a specific and easy detected gene marker of ansamycin producing potential of actinomycetes Multi alignment of know n AHBA synthases involved in ansamycin biosynthesis indicated that AHBA synthases are highly con- served. In this paper, a pair of oligonucleotides w as designed successfully for PCR amplification/ detection specifically of the 755bp portion of the AHBA synthase genes from unspecified actinomycetes strains U sing. , - this pair of oligonucleotides a high throughput PCR screening method for potential ansamycin producers from actinomycetes strains w as established. Of the 1900 unspecified soil-isolated strains screened, 33 strains con- tained AHBA synthase gene. T hese strains w ere likely to be potential ansamycin producing strains. KEY WORDSAnsamycins; AHBA synthase gene; Polymerase chain reaction ( PCR) ; Molecular screening; Actinomycetes