Two pHZ1358-derivative vectors for efficient gene knockout in streptomyces.
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.
He
Yunlong
Y
Laboratory of Microbiol Metabolism, Shanghai Jiaotong University, Shanghai. China.
Wang
Zhijun
Z
Bai
Linquan
L
Liang
Jingdan
J
Zhou
Xiufen
X
Deng
Zixin
Z
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*
D004269:DNA, Bacterial; D005819:Genetic Markers