Kudriavtseva E A, Yun Yang L, Voeĭkova T A, Lomovskaia N D
Genetika. 1994 Jul;30(7):886-97.
Opportunities for application of integrative vectors carrying the attP site and the int gene of the temperate actinophage phi C31 for cloning genes in Streptomyces strains were demonstrated. The behavior of the integrative vectors pZAT22 and pTO1 in the model strain S. lividans TK64 and in the bialaphos-producing strain S. hygroscopicus, respectively, was characterized. Restriction maps of the S. lividans and S. hygroscopicus chromosomal regions containing attB sites were constructed. The bar gene of resistance to bialaphos was incorporated into the chromosome of the model strain S. lividans via the integrative pZAT22 vector, the level of expression of this gene within the chromosome of a heterologous host was determined. The possibility of amplification of the bar gene in the chromosome of the strain S. lividans was shown. The conjugative integrative vector pTO1, which carried the cloned bar gene, was introduced into the bialaphos-producing strain by intergeneric matings of Escherichia coli and S. hygroscopicus. The effect of an additional copy of the gene in the chromosome of S. hygroscopicus on strain resistance and productivity was studied.
证明了携带温和型放线菌噬菌体phi C31的attP位点和int基因的整合载体在链霉菌菌株中克隆基因的应用机会。分别对整合载体pZAT22和pTO1在模式菌株变铅青链霉菌TK64和产双丙氨膦的吸水链霉菌中的行为进行了表征。构建了含有attB位点的变铅青链霉菌和吸水链霉菌染色体区域的限制性图谱。通过整合型pZAT22载体将抗双丙氨膦的bar基因整合到模式菌株变铅青链霉菌的染色体中，测定了该基因在异源宿主染色体中的表达水平。显示了变铅青链霉菌染色体中bar基因扩增的可能性。通过大肠杆菌和吸水链霉菌的属间交配，将携带克隆bar基因的接合整合载体pTO1导入产双丙氨膦的菌株中。研究了吸水链霉菌染色体中该基因的额外拷贝对菌株抗性和生产力的影响。
