Development of Series of Affinity Tags in Streptomyces.
Development of Series of Affinity Tags in Streptomyces.
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亲和系列的开发
DOI：
10.1038/s41598-017-07377-4
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发表时间：
2017-07-31
影响因子：
4.6
通讯作者：
Li YQ
中科院分区：
文献类型：
--
作者：
Mao XM;Sun N;Zheng Y;Li YQ
Streptomycesare of great biological and industrial significance due to their complex morphological development and ability to produce numerous secondary metabolites. However, the intrinsic biochemical mechanisms underlying morphogenesis and secondary metabolism are rarely revealed, partially because of the limited availability of the biochemical tools inStreptomyces. Here we provided series of integrative vectors with various affinity tags, including single tags 3×FLAG, 3×HA, 3×Strep-tag II, 18×His, 13×Myc, and dual tags, all of which were driven from a strong constitutive promoterermEp*. Using a sigma factor SigT fromS. coelicoloras a model, we successfully expressed and immuno-detected SigT fused with all tags. Moreover, after SigT was N-terminally tagged with 3×FLAG and C-terminally tagged with 18×His, we isolated SigT-interactive proteins from theS. coelicolorlysate based on the tandem affinity purification (TAP). Particularly, among the proteins purified, the SigT cognate anti-sigma factor RstA ranked the top with the most total independent spectra. These data suggested the feasibility of these affinity tags inStreptomyces, which will be widely employed to explore the biochemical mechanisms to further understand the dynamic and elaborate regulation in this genus.
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影响因子：
11.3
作者：
McCormick JR;Flärdh K
通讯作者：
Flärdh K
影响因子：
14.8
作者：
Schmidt, Thomas G. M.;Skerra, Arne
通讯作者：
Skerra, Arne
影响因子：
4.3
作者：
Richards DM;Hempel AM;Flärdh K;Buttner MJ;Howard M
通讯作者：
Howard M
影响因子：
14.9
作者：
Sola-Landa A;Rodríguez-García A;Amin R;Wohlleben W;Martín JF
通讯作者：
Martín JF
影响因子：
5.8
作者：
Donczew M;Mackiewicz P;Wróbel A;Flärdh K;Zakrzewska-Czerwińska J;Jakimowicz D
通讯作者：
Jakimowicz D