Expression of membrane proteins from Mycobacterium tuberculosis in Escherichia coli as fusions with maltose binding protein
Expression of membrane proteins from Mycobacterium tuberculosis in Escherichia coli as fusions with maltose binding protein
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DOI：
10.1016/j.pep.2006.11.022
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发表时间：
2007-05-01
影响因子：
1.6
通讯作者：
Gao, F.
中科院分区：
文献类型：
--
作者：
Korepanova, A.;Moore, J. D.;Gao, F.
来自结核分枝杆菌的 22 种低分子量整合膜蛋白中的 16 种先前表达水平较低或检测不到，在大肠杆菌中表达为与麦芽糖结合蛋白 (MBP) 和 Hiss 标签的融合体。 68% 的目标蛋白以可溶性和/或包涵体形式高产量 (> 30 mg/L) 表达。对于表达为可溶性蛋白的 13 种蛋白中的 10 种和仅表达为包涵体的三种蛋白，凝血酶成功切割 MBP 融合蛋白。与 Luria-Bertani (LB) 生长培养基相比，使用自诱导生长培养基提高了 75% 表达蛋白的产量。事实证明，在将麦芽糖结合蛋白作为融合标签附着后，从一组先前低表达和非表达的蛋白质中表达出适合结构研究的产量的整合膜蛋白是非常成功的。 (c) 2006 Elsevier Inc. 保留所有权利。
Sixteen of 22 low molecular weight integral membrane proteins from Mycobacterium tuberculosis with previously poor or undetectable levels of expression were expressed in Escherichia coli as fusions with both the maltose binding protein (MBP) and a Hiss-tag. Sixty-eight percent of targeted proteins were expressed in high yield (> 30 mg/L) in soluble and/or inclusion body form. Thrombin cleavage of the MBP fusion protein was successful for 10 of 13 proteins expressed as soluble proteins and for three proteins expressed only as inclusion bodies. The use of autoinduction growth media increased yields over Luria-Bertani (LB) growth media in 75% of the expressed proteins. Expressing integral membrane proteins with yields suitable for structural studies from a set of previously low and non-expressing proteins proved highly successful upon attachment of the maltose binding protein as a fusion tag. (c) 2006 Elsevier Inc. All rights reserved.