A gene cloning system for the siomycin producer Streptomyces sioyaensis NRRL-B5408
A gene cloning system for the siomycin producer Streptomyces sioyaensis NRRL-B5408
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DOI：
10.1007/s12223-009-0013-x
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发表时间：
2009-05
影响因子：
2.6
通讯作者：
M. Myronovskyy;B. Ostash;I. Ostash;V. Fedorenko
中科院分区：
文献类型：
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作者：
M. Myronovskyy;B. Ostash;I. Ostash;V. Fedorenko
Sioyaensis链霉菌NRRL-B5408产生一种硫霉素复合物（一组结构上与硫链丝菌素相关的硫肽抗生素）。描述了用于检测硅霉素生产和DNA转移到该菌株中的遗传工具的开发。对现有的基于tipA的报告基因系统进行了改造，以实现其稳定整合到放线菌基因组中。将各种复制型质粒（pKC 1139、pKC 1218 E、pSOK 101）以及基于放线菌噬菌体C31和VWB的载体pSET 152和pSOK 804分别弗罗姆.大肠杆菌进入硫霉素产生菌的频率范围为3.7 × 10 - 9 ~ 1.1 × 10 - 5。接合子在产生硫霉素的能力上与野生型没有区别。每个整合质粒有一个attB位点。通过破坏推定的非核糖体肽合成酶基因，证实了pKC 1139的温度敏感复制子在S. sioyaensisinc中用于插入基因失活的效用。
Streptomyces sioyaensisNRRL-B5408 produces a siomycin complex (a group of thiopeptide antibiotics structurally related to thiostrepton). Development of genetic tools for the detection of siomycin production and DNA transfer into this strain is described. The existingtipA-based reporter system for determination of siomycin production was modified to achieve its stable integration into actinomycete genomes. Various replicative plasmids (pKC1139, pKC1218E, pSOK101) as well as actinophage ϕC31- and VWB-based vectors pSET152 and pSOK804, respectively, were conjugally transferred fromE. coliinto the siomycin producer at a frequency ranging from 3.7 × 10−9to 1.1 × 10−5. The transconjugants did not differ from wild type in their ability to produce siomycin. There is oneattBsite for each integrative plasmid. The utility of temperature sensitive replicon of pKC1139 for insertional gene inactivation inS. sioyaensishas been validated by disruption of putative nonribosomal peptide synthetase gene.