Soldatova L I, Sladkova I A, Orekhov A V
Antibiot Khimioter. 1994 Jun;39(6):3-7.
Actinomycete integrative vectors were constructed. The vectors contain the Escherichia coli plasmid ColE1 replicon, the thiostrepton resistance gene used for selection in Streptomyces and a fragment of the phi C31 actinophage genome with integrative functions. The pS133 and PS135 vector DNAs transformed Streptomyces bambergiensis 712, a strain producing the phosphoglycolipid antibiotic moenomycin. Two types of the transformants were detected. The first type was not affected in the ability to produce moenomycin and the vector pS135 DNA was shown to integrate into the S. bambergiensis 712 genome by the site-specific pattern with the psi C31 phage DNA fragment. The second type of the transformants lost the ability to produce moenomycin. The Southern analysis and cloning of the inserted DNA indicated that in this case the vector pS135 and pS133 DNAs also integrated specifically into the genome but the integration took place not within the phage DNA fragment. Its realization was suggested to proceed via homologous recombination.
构建了放线菌整合载体。这些载体包含大肠杆菌质粒ColE1复制子、用于在链霉菌中筛选的硫链丝菌素抗性基因以及具有整合功能的phi C31放线噬菌体基因组片段。pS133和PS135载体DNA转化了产磷酸糖脂抗生素莫能菌素的菌株巴氏链霉菌712。检测到两种类型的转化体。第一种类型在产生莫能菌素的能力上没有受到影响，并且载体pS135 DNA通过与psi C31噬菌体DNA片段的位点特异性模式整合到巴氏链霉菌712基因组中。第二种类型的转化体失去了产生莫能菌素的能力。对插入DNA的Southern分析和克隆表明，在这种情况下，载体pS135和pS133 DNA也特异性地整合到基因组中，但整合不是发生在噬菌体DNA片段内。推测其实现是通过同源重组进行的。
