Development of an intergeneric conjugal transfer system for xinaomycins-producing Streptomyces noursei xinao-4

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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 \r<br />of conjugation (8 × 10−3 exconjugants per recipient) was obtained when spores of \r<br />S. noursei xinao-4 were heat-shocked at 50 °C for 10 min, mixed with Escherichia coli \r<br />ET12567 (pUZ8002/pSET152) in the ratio of 1:100, plated on 2CMY medium \r<br />containing 40 mmol/L MgCl2, and incubated at 30 °C for 22 h. With this protocol, \r<br />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 \r<br />to validate the above intergeneric conjugal transfer system, a glycosyltransferase gene \r<br />was cloned and efficiently knocked out in S. noursei xinao-4 using pSG5-based \r<br />plasmid pKC1139.

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Sun, F. H., Luo, D., Shu, D., Zhong, J., & Tan, H. (2014). Development of an intergeneric conjugal transfer system for xinaomycins-producing Streptomyces noursei xinao-4. International Journal of Molecular Sciences, 15(7), 12217–12230. https://doi.org/10.3390/ijms150712217

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