Genome engineering and direct cloning of antibiotic gene clusters via phage ϕBT1 integrase-mediated site-specific recombination in Streptomyces.
Genome engineering and direct cloning of antibiotic gene clusters via phage ϕBT1 integrase-mediated site-specific recombination in Streptomyces.
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通过噬菌体ϕBT1整合酶介导的链霉菌位点特异性重组进行基因组工程和抗生素基因簇的直接克隆。
DOI：
10.1038/srep08740
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发表时间：
2015-03-04
影响因子：
4.6
通讯作者：
Niu G
中科院分区：
文献类型：
--
作者：
Du D;Wang L;Tian Y;Liu H;Tan H;Niu G
已经使用了几种策略直接从细菌基因组克隆大的DNA片段。这些方法中的大多数是基于由专门的重组酶及其靶位点组成的不同位点特异性重组系统。本研究开发了一种基于噬菌体NTBT1整合酶介导的位点特异性重组的新策略，并用于链霉菌基因组工程和抗生素基因簇的同步克隆。该方法成功地克隆了天蓝色链霉菌M145的放线菌紫素基因簇、玫瑰孢链霉菌NRRL 15998的萘莎霉素基因簇和达托霉素基因簇，克隆频率均在80%以上。此外，该系统可用于提高抗生素的效价，如我们所证明的放线菌紫素和达托霉素，它将广泛适用于许多链霉菌。
Several strategies have been used to clone large DNA fragments directly from bacterial genome. Most of these approaches are based on different site-specific recombination systems consisting of a specialized recombinase and its target sites. In this study, a novel strategy based on phage ϕBT1 integrase-mediated site-specific recombination was developed, and used for simultaneous Streptomyces genome engineering and cloning of antibiotic gene clusters. This method has been proved successful for the cloning of actinorhodin gene cluster from Streptomyces coelicolor M145, napsamycin gene cluster and daptomycin gene cluster from Streptomyces roseosporus NRRL 15998 at a frequency higher than 80%. Furthermore, the system could be used to increase the titer of antibiotics as we demonstrated with actinorhodin and daptomycin, and it will be broadly applicable in many Streptomyces.
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影响因子：
4.6
作者：
Fernández-Martínez LT;Bibb MJ
通讯作者：
Bibb MJ
影响因子：
3.2
作者：
Kaysser, Leonard;Tang, Xiaoyu;Gust, Bertolt
通讯作者：
Gust, Bertolt
影响因子：
4.4
作者：
Herrmann, Simone;Siegl, Theresa;Luzhetskyy, Andriy
通讯作者：
Luzhetskyy, Andriy
影响因子：
4.4
作者：
Kvitko, Brian H.;McMillan, Ian A.;Schweizer, Herbert P.
通讯作者：
Schweizer, Herbert P.
DOI：
10.1073/pnas.0337542100
发表时间：
2003-02-18
影响因子：
11.1
作者：
Gust, B;Challis, GL;Chater, KF
通讯作者：
Chater, KF