Journal of Microbiology and Biotechnology
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2010 ; 20(4): 678~682

AuthorYunlong He, Zhijun Wang, Linquan Bai, Jingdan Liang, Xiufen Zhou, Zixin Deng
TitleTwo pHZ1358-derivative Vectors for Efficient Gene Knockout in Streptomyces
PublicationInfo J. Microbiol. Biotechnol.2010 ; 20(4): 678~682
AbstractDeletion 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.
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Keywordsshuttle vector, pHZ1358, pJTU1278, pJTU1289, conjugation, PCR-targeting
References
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