Chong Pei Pei, Podmore Sylvia M, Kieser Helen M, Redenbach Matthias, Turgay Kürsad, Marahiel Mohamed, Hopwood David A, Smith Colin P
Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology (UMIST), PO Box 88, Manchester M60 1QD, UK.
John Innes Centre, Norwich NR4 7UH, UK.
Microbiology (Reading). 1998 Jan;144 ( Pt 1):193-199. doi: 10.1099/00221287-144-1-193.
Putative peptide-synthetase-encoding DNA fragments were isolated from the Streptomyces coelicolor A3(2) chromosome using a PCR-based approach and mapped to a single approximately 35 kb segment. In integrative transformation experiments, DNA fragments from this region disrupted production of the calcium-dependent antibiotic (CDA) and had sequences characteristic of non-ribosomal peptide synthetases, thus proving that the cda locus had been cloned.
采用基于聚合酶链式反应(PCR)的方法,从天蓝色链霉菌A3(2)染色体中分离出推测的肽合成酶编码DNA片段,并将其定位到一个约35 kb的单一区段。在整合转化实验中,该区域的DNA片段破坏了钙依赖性抗生素(CDA)的产生,并且具有非核糖体肽合成酶的序列特征,从而证明已克隆到cda基因座。