链霉菌中抗生素抗性基因的克隆

Cloning of antibiotic-resistance genes in Streptomyces.

作者信息

Murakami T, Nojiri C, Toyama H, Hayashi E, Katumata K, Anzai H, Matsuhashi Y, Yamada Y, Nagaoka K

出版信息

J Antibiot (Tokyo). 1983 Oct;36(10):1305-11. doi: 10.7164/antibiotics.36.1305.

DOI:10.7164/antibiotics.36.1305

Abstract

Antibiotic-resistance genes were shotgun cloned from antibiotic-producing Streptomyces sp. using pock-forming plasmids (pSF689 and pSF765), as cloning vectors. Streptomyces chartreusis SF1623 and S. lividans 66 were used as host strains. The ribostamycin (RSM) resistance gene was cloned from S. ribosidificus SF733 DNA (on a 2.3 Md PstI fragment) into both S. chartreusis SF1623 and S. lividans 66, using pSF689 as vector. Kanamycin (KM), novobiocin (NB), destomycin (DM) and racemomycin (RM) resistance genes were cloned from S. kanamyceticus M1164, S. spheroides M1469, S. rimofaciens M1470 and S. lavendulae A249 genomic DNA into S. lividans 66, using pSF765 as vector. Furthermore two types of KM resistance determinants derived from S. kanamyceticus M1164 were cloned using S. lividans 66, the pSF689 vector. The RSM resistance gene showed no homology to plasmid pSF733 of S. ribosidificus SF733, but hybridized to PstI or BclI digested total DNA of S. ribosidificus SF733.

摘要

利用噬菌斑形成质粒（pSF689和pSF765）作为克隆载体，从产抗生素的链霉菌属中鸟枪法克隆抗生素抗性基因。将浅绿链霉菌SF1623和变铅青链霉菌66用作宿主菌株。使用pSF689作为载体，将核糖霉素（RSM）抗性基因从核糖苷链霉菌SF733 DNA（位于一个2.3 Md的PstI片段上）克隆到浅绿链霉菌SF1623和变铅青链霉菌66中。使用pSF765作为载体，将卡那霉素（KM）、新生霉素（NB）、龟裂链霉素（DM）和消旋霉素（RM）抗性基因从卡那霉素链霉菌M1164、球孢链霉菌M1469、边缘链霉菌M1470和薰衣草链霉菌A249基因组DNA克隆到变铅青链霉菌66中。此外，使用变铅青链霉菌66和pSF689载体克隆了源自卡那霉素链霉菌M1164的两种类型的KM抗性决定簇。RSM抗性基因与核糖苷链霉菌SF733的质粒pSF733没有同源性，但与核糖苷链霉菌SF733经PstI或BclI消化的总DNA杂交。