Open AccessRecombinational cloning of the antibiotic biosynthetic gene clusters in linear plasmid SCP 1 of Streptomyces coelicolor A3(2)
Author(s) -
Zhang Ran,
Xia Haiyang,
Xu Qingyu,
Dang Fujun,
Qin Zhongjun
Publication year - 2013
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/1574-6968.12183
Subject(s) - streptomyces coelicolor , cosmid , plasmid , biology , genetics , cloning (programming) , gene , streptomyces , molecular cloning , dna , homologous recombination , microbiology and biotechnology , gene expression , bacteria , computer science , programming language
The model organism Streptomyces coelicolor A3(2) harbors a 356‐kb linear plasmid, SCP1. We report here development of a recombinational cloning method for deleting large segment from one telomere of SCP1 followed by replacing with the telomere of pSLA 2 and sequentially inserting with the overlapping cosmids in vivo . The procedure depends on homologous recombination coupled with cleavage at telomere termini by telomere terminal protein. Using this procedure, we cloned the 81‐kb avermectin and the 76‐kb spinosad biosynthetic gene clusters into SCP1. Heterologous expression of avermectin production in S. coelicolor was detected. These results demonstrate the utility of SCP1 for cloning large DNA segments such as antibiotic biosynthetic gene clusters.