title: Two pHZ1358-derivative vectors for efficient gene knockout in streptomyces.
abstract: Deletion of sti on Streptomyces plasmid pIJ101, made its derivative pHZ1358 an efficient vector for gene disruption and replacement. Here, we further optimized pHZ1358 by the construction of a derivative plasmid pJTU1278, in which a cassette carrying multiple cloning sites and a lacZ selection marker was introduced for convenient plasmid construction in E. coli. Furthermore, we also deleted oriT region of the pJTU1278, generated a vector (pJTU1289) that can be used specifically for PCR-targeting. The efficient usage of these vectors was demonstrated by the deletion of a gene involved in avermectin biosynthetic in S. avermitilis.
authors.affiliation: Laboratory of Microbiol Metabolism, Shanghai Jiaotong University, Shanghai. China.
authors.forename: Yunlong
authors.initials: Y
authors.lastname: He
authors.forename: Zhijun
authors.initials: Z
authors.lastname: Wang
authors.forename: Linquan
authors.initials: L
authors.lastname: Bai
authors.forename: Jingdan
authors.initials: J
authors.lastname: Liang
authors.forename: Xiufen
authors.initials: X
authors.lastname: Zhou
authors.forename: Zixin
authors.initials: Z
authors.lastname: Deng
mesh_terms: D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D055786:Gene Knockout Techniques / Q000379:methods*; D005819:Genetic Markers; D005822:Genetic Vectors / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013302:Streptomyces / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D005819:Genetic Markers