The cloned Streptomyces bikiniensis A-factor determinant.
The cloned Streptomyces bikiniensis A-factor determinant.
复制标题
克隆的比基尼链霉菌 A 因子决定簇。
DOI：
10.7164/antibiotics.38.636
复制
发表时间：
1985
期刊：
影响因子：
--
通讯作者：
T. Beppu
中科院分区：
文献类型：
--
作者：
S. Horinouchi;M. Nishiyama;H. Suzuki;Y. Kumada;T. Beppu
利用限制性内切酶对产链霉素链霉菌（Streptomyces bikiniensis, Horinouchi et al.， J. Bacteriol. 158: 481约487,1984）的a因子决定因子片段进行了克隆和鉴定。携带1.1 kb片段的杂交多拷贝质粒（pAFB15）使S. bikininiensis和Streptomyces griseus缺乏A因子的突变体以及Streptomyces colicolor A3的afsA突变体大量产生A因子(2)。利用启动子-探针载体pARC 1鉴定了1.1 kb片段中的转录控制信号。质粒pAFB15在4株原不能产生a因子的链霉菌中也能产生a因子，且具有明显的基因剂量效应，说明a因子的前体是链霉菌中常见的代谢物，仅获得1.1 kb片段编码的一个关键酶就足以使链霉菌合成a因子。
By cleavage with restriction endonucleases and cloning the resultant fragments, the A-factor determinant cloned from streptomycin-producing Streptomyces bikiniensis (Horinouchi et al., J. Bacteriol. 158: 481 approximately 487, 1984) was narrowed down to a 1.1-kilobase (kb) fragment. A hybrid multicopy plasmid (pAFB15) carrying the 1.1-kb fragment conferred A-factor production in a large quantity to A-factor-deficient mutants of S. bikiniensis and Streptomyces griseus as well as afsA mutants of Streptomyces coelicolor A3(2). A transcriptional control signal in the 1.1-kb fragment was identified by using a promoter-probe vector pARC 1. Plasmid pAFB15 also caused A-factor production with a marked gene dosage effect in four different Streptomyces strains which originally had no ability to produce A-factor and no DNA sequence homologous to the S. bikiniensis A-factor determinant, suggesting that precursors of A-factor are common metabolites in streptomycetes and that acquisition of only a single key enzyme encoded by the 1.1-kb fragment is sufficient for Streptomyces strains to synthesize A-factor.