期刊
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
卷 20, 期 4, 页码 678-682出版社
KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
DOI: 10.4014/jmb.0910.10031
关键词
Shuttle vector; pHZ1358; pJTU1278; pJTU1289; conjugation; PCR-targeting
资金
- National Science Foundation of China
- Ministry of Science and Technology
- Ministry of Education of China
- Shanghai Municipal Council of Science and Technology and Shanghai Leading Academic Discipline
- State Key Laboratory of Bio-organic and Natural Products Chemistry (CAS)
The deletion of sti from the Streptomyces plasmid pIJ101 made its derivative pHZ1358 an efficient vector for gene disruption and replacement. Here, pHZ1358 was further optimized by the construction of a derivative plasmid pJTU1278, in which a cassette carrying multiple cloning sites and a lacZ selection marker were introduced for convenient plasmid construction in E. coli. In addition, the oriT region of pJTU1278 was also deleted, generating a vector (pJTU1289) that can be used specifically for PCR-targeting. The efficient usage of these vectors was demonstrated by the deletion of the gene involved in avermectin biosynthesis in S. avermitilis.
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