Starodubtseva L I, Danilenko V N
Mol Biol (Mosk). 1989 Jan-Feb;23(1):193-203.
The S. rimosus amplifying sequence AUD-Sr1 encodes kanamycin and neomycin resistance, defined in the case of neomycin by aminoglycoside phosphotransferase. Its cloning on plasmid SLP1.2 makes possible the co-amplification of the obtained hybrid plasmids in S. lividans. In our study the regions responsible for resistance to aminoglycoside antibiotics and the capacity for amplification the two hybrid plasmids pSU10 and pSU3 were determined. Experiments on subcloning of the AUD-Sr1 sequence fragments on vector pIJ702 revealed localization of kanamycin and neomycin resistance determinants between PvuII(6) and BglII(7) on the AUD-Sr1 sequence fragments of 2.0 kb length. Two regions responsible for amplification of the hybrid plasmids were detected with deletion and insertion mapping. The first region is localized in the region of the plasmid SLP1.2 BamHI site and the second region is localized on the PstI(4)-PvuII(6) of the AUD-Sr1 sequence fragment of 1.1 kb length.
龟裂链霉菌扩增序列AUD-Sr1编码卡那霉素和新霉素抗性,就新霉素而言,这种抗性由氨基糖苷磷酸转移酶定义。它在质粒SLP1.2上的克隆使得所得杂交质粒在变铅青链霉菌中能够共同扩增。在我们的研究中,确定了对氨基糖苷类抗生素的抗性区域以及两种杂交质粒pSU10和pSU3的扩增能力。将AUD-Sr1序列片段亚克隆到载体pIJ702上的实验表明,卡那霉素和新霉素抗性决定簇位于长度为2.0 kb的AUD-Sr1序列片段上的PvuII(6)和BglII(7)之间。通过缺失和插入图谱检测到两个负责杂交质粒扩增的区域。第一个区域位于质粒SLP1.2 BamHI位点区域,第二个区域位于长度为1.1 kb的AUD-Sr1序列片段的PstI(4)-PvuII(6)上。