委内瑞拉链霉菌ISP 5230邻氨基苯甲酸合成酶基因的分子克隆
Molecular cloning of the genes for anthranilate synthetase from Streptomyces venezuelae ISP 5230.
作者信息
Paradkar A S, Stuttard C, Vining L C
机构信息
Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada.
出版信息
FEMS Microbiol Lett. 1991 Mar 1;62(2-3):177-81. doi: 10.1016/0378-1097(91)90154-3.
Fragments of genomic DNA from Streptomyces venezuelae ISP5230 were cloned in the Escherichia coli expression vector pTZ18R and the plasmids were used to transform E. coli JA194 (trpE). The transformants included a prototrophic strain containing a recombinant plasmid, pDQ181, with an approximately 6.8-kb insert. Subcloning located the trpE-complementing DNA in a 2.4-kb segment. Transformation of E. coli ED23 (lacking both trpE and trpG functions) with plasmids containing the 2.4-kb DNA segment gave prototrophic strains exhibiting both the ASI and ASII activities of anthranilate synthetase. The results indicated that trpE and trpG are clustered in S. venezuelae. Regions hybridizing to the pDQ181 insert were present in the genomic DNA of other streptomycetes.
委内瑞拉链霉菌ISP5230的基因组DNA片段被克隆到大肠杆菌表达载体pTZ18R中，这些质粒被用于转化大肠杆菌JA194（trpE）。转化子包括一个含有重组质粒pDQ181的原养型菌株，该质粒带有一个约6.8 kb的插入片段。亚克隆将trpE互补DNA定位在一个2.4 kb的片段中。用含有2.4 kb DNA片段的质粒转化大肠杆菌ED23（缺乏trpE和trpG功能），得到了表现出邻氨基苯甲酸合成酶ASI和ASII活性的原养型菌株。结果表明，trpE和trpG在委内瑞拉链霉菌中是成簇的。与pDQ181插入片段杂交的区域存在于其他链霉菌的基因组DNA中。
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