Development of an Intergeneric Conjugal Transfer System for Xinaomycins-Producing Streptomyces noursei Xinao-4 |
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| Authors: | Feng-Hui Sun Di Luo Dan Shu Juan Zhong Hong Tan |
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| Affiliation: | 1.Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, No. 9 Section 4, Renmin Nan Road, Chengdu 610041, China; E-Mails: (F.-H.S.); (D.L.); (D.S.); (J.Z.);2.Environmental Microbiology Key Laboratory of Sichuan Province, No. 9 Section 4, Renmin Nan Road, Chengdu 610041, China;3.University of the Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing 100049, China |
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| Abstract: | To introduce DNA into Streptomyces noursei xinao-4, which produces xinaomycins, we explored an intergeneric conjugal transfer system. High efficiency of conjugation (8 × 10−3 exconjugants per recipient) was obtained when spores of S. noursei xinao-4 were heat-shocked at 50 °C for 10 min, mixed with Escherichia coli ET12567 (pUZ8002/pSET152) in the ratio of 1:100, plated on 2CMY medium containing 40 mmol/L MgCl2, and incubated at 30 °C for 22 h. With this protocol, the plasmids pKC1139 and pSET152 were successfully transferred from E. coli ET12567 (pUZ8002) with different frequencies. Among all parameters, the ratio of donor to recipient cell number had the strongest effect on the transformation efficiency. In order to validate the above intergeneric conjugal transfer system, a glycosyltransferase gene was cloned and efficiently knocked out in S. noursei xinao-4 using pSG5-based plasmid pKC1139. |
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| Keywords: | Streptomyces noursei xinaomycins intergeneric conjugation glycosyltransferase |
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