Huan L D, Dong K N, Zhuang Z H, Xue Y G
Institute of Microbiology, Academia Sinica, Beijing.
Yi Chuan Xue Bao. 1991;18(1):82-9.
BamHI restriction fragments from Streptomyces lividans TK24 chromosome DNA have been cloned into BamHI site of promoter probe plasmid pIJ486. Transformants were selected on the medium containing 5 micrograms/ml of neomycin. Four recombinant plasmids pMG1(10.6 kb), pMG40(7.6 kb), pMG50(10.8 kb) and pMG88(7.92 kb), were found and designated respectively. The inserted fragments in pMG40 and pMG50 were reduced to 0.78kb and 2.2 kb by BglII digestion and rejoining. The different levels of neomycin and kanamycin resistance of these recombinant plasmids were determined. The results revealed that pMG50-25 showed a high level of neomycin resistance (90 micrograms/ml) and kanamycin resistance (500 micrograms/ml).
来自变铅青链霉菌TK24染色体DNA的BamHI限制性片段已被克隆到启动子探针质粒pIJ486的BamHI位点。在含有5微克/毫升新霉素的培养基上筛选转化体。发现并分别命名了四个重组质粒pMG1（10.6 kb）、pMG40（7.6 kb）、pMG50（10.8 kb）和pMG88（7.92 kb）。通过BglII消化和重新连接，pMG40和pMG50中的插入片段分别减少到0.78 kb和2.2 kb。测定了这些重组质粒对新霉素和卡那霉素的不同抗性水平。结果表明，pMG50-25表现出高水平的新霉素抗性（90微克/毫升）和卡那霉素抗性（500微克/毫升）。
