Kuhstoss S, Richardson M A, Rao R N
Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, IN 46285.
Gene. 1991 Jan 2;97(1):143-6. doi: 10.1016/0378-1119(91)90022-4.
Cloning vectors based on the Streptomyces ambofaciens plasmid pSAM2 and the streptomycete phage phi C31 were developed for use in Streptomyces spp. These vectors replicate in Escherichia coli but integrate by site-specific recombination in Streptomyces spp. Both pSAM2-based and phi C31-based vectors transformed a number of different Streptomyces spp; however, the phi C31-based vectors consistently transformed at higher frequencies than pSAM2-based vectors. Southern analysis indicated that the phi C31-based vectors integrated at a unique site in the S. ambofaciens chromosome, while the pSAM2-based vectors gave complex patterns which could indicate structural instability or use of multiple loci. Both types of vectors utilize the apramycin (Am)-resistance gene which can be selected in E. coli and Streptomyces spp. with either Am or the commercially available antibiotic Geneticin (G418).
基于浅青紫链霉菌质粒pSAM2和链霉菌噬菌体phi C31构建的克隆载体被开发用于链霉菌属。这些载体在大肠杆菌中复制，但通过位点特异性重组整合到链霉菌属中。基于pSAM2和基于phi C31的载体都能转化多种不同的链霉菌属；然而，基于phi C31的载体转化频率始终高于基于pSAM2的载体。Southern分析表明，基于phi C31的载体整合到浅青紫链霉菌染色体的一个特定位点，而基于pSAM2的载体产生复杂的模式，这可能表明结构不稳定或使用了多个位点。两种类型的载体都利用了阿泊拉霉素（Am）抗性基因，该基因可在大肠杆菌和链霉菌属中用Am或市售抗生素遗传霉素（G418）进行选择。
