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构建一种新型的依赖双精氨酸转运(Tat)的表达载体,用于在谷氨酸棒杆菌中分泌生产异源蛋白。

Construction of a novel twin-arginine translocation (Tat)-dependent type expression vector for secretory production of heterologous proteins in Corynebacterium glutamicum.

作者信息

Zhang Lirong, Jia Huimin, Xu Daqing

机构信息

College of Plant Protection, Agricultural University of Hebei, Baoding 071001, China.

College of Life Sciences, Agricultural University of Hebei, Baoding 071001, China.

出版信息

Plasmid. 2015 Nov;82:50-5. doi: 10.1016/j.plasmid.2015.10.004. Epub 2015 Oct 22.

Abstract

Corynebacterium glutamicum is recognized as a favorable host for the secretory production of heterologous proteins. However, there are few secretion-type expression vectors available for protein production in C. glutamicum. In this study, we constructed a shuttle expression vector pAU3, which harbors the strong promoter tac-M for constitutive gene transcription, the consensus RBS sequence for protein translation, and the strong cgR_0949 signal sequence for protein secretion via the Tat pathway in C. glutamicum. The applicability of pAU3 was confirmed by the highly efficient expression and secretion of the CAT protein in C. glutamicum. The vector pAU3 is highly useful for secretory production of heterologous proteins in C. glutamicum.

摘要

谷氨酸棒杆菌被认为是分泌生产异源蛋白的理想宿主。然而,可用于谷氨酸棒杆菌中蛋白质生产的分泌型表达载体很少。在本研究中,我们构建了穿梭表达载体pAU3,其含有用于组成型基因转录的强启动子tac-M、用于蛋白质翻译的共有核糖体结合位点(RBS)序列以及用于通过谷氨酸棒杆菌中的Tat途径进行蛋白质分泌的强cgR_0949信号序列。通过在谷氨酸棒杆菌中高效表达和分泌CAT蛋白证实了pAU3的适用性。载体pAU3对于在谷氨酸棒杆菌中分泌生产异源蛋白非常有用。

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