Grigor'ev A E, Zhukov D B, Orlova V A, Danilenko V N
Antibiot Khimioter. 1991 Nov;36(11):3-5.
The integrative vectors pSU 475 and pSU 476 with variable numbers of copies per genome were developed for antibiotic producing actinomycetes. For this, the amplifying sequence AUD-Sr 1 of Streptomyces rimosus and the BamHIB fragment of the eSA 1 genetic element from Streptomyces antibioticus were used. The eSA 1 fragment was an element required for integration of a vector to the actinomycete chromosomes since it was homologous with the chromosomal DNAs of S. lividans, S. erythraeus and S. antibioticus. At the first stage the AUD-Sr 1 sequence within the actinomycete plastid pSU 23 was cloned by the vector pUC 19 to E coli. In that experiment the 12.4-kb plasmid pSU 449 was isolated. At the second stage the BamHIB-fragment of the eSA 1 element was incorporated into the resultant hybrid plasmid pSU 449. The 16.5-kb hybrid plasmids pSU 475 and pSU 476 were isolated. In these plasmids the BamHIB fragment of eSA 1 was present in two orientations. The developed vectors were useful in cloning DNA to S. lividans and S. erythraeus.
为抗生素生产放线菌开发了每个基因组具有可变拷贝数的整合载体pSU 475和pSU 476。为此，使用了龟裂链霉菌的扩增序列AUD-Sr 1和来自抗生链霉菌的eSA 1遗传元件的BamHIB片段。eSA 1片段是载体整合到放线菌染色体所需的元件，因为它与变铅青链霉菌、红霉素链霉菌和抗生链霉菌的染色体DNA同源。在第一阶段，通过载体pUC 19将放线菌质体pSU 23内的AUD-Sr 1序列克隆到大肠杆菌中。在该实验中，分离出了12.4 kb的质粒pSU 449。在第二阶段，将eSA 1元件的BamHIB片段整合到所得的杂交质粒pSU 449中。分离出了16.5 kb的杂交质粒pSU 475和pSU 476。在这些质粒中，eSA 1的BamHIB片段以两种方向存在。所开发的载体可用于将DNA克隆到变铅青链霉菌和红霉素链霉菌中。
