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pHsh 载体,一种新型大肠杆菌表达系统,用于大规模生产重组酶。

pHsh vectors, a novel expression system of Escherichia coli for the large-scale production of recombinant enzymes.

机构信息

Jiangsu Key Laboratory for Biodiversity and Bioresources, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing, 210046, People's Republic of China.

出版信息

Biotechnol Lett. 2010 Jun;32(6):795-801. doi: 10.1007/s10529-010-0223-y. Epub 2010 Feb 14.

Abstract

Gene expression system Hsh is developed to increase enzyme production and to decrease the cost in the induction of gene expression in Escherichia coli. The vectors of Hsh system were constructed by combining a synthesized heat-shock promoter with a synthesized terminator and an origin of replication derived from pUC19 in which the expression of foreign genes was regulated by an alternative sigma factor, sigma(32) of E. coli. In comparison, the Hsh promoter gave a 2.4-fold higher production for xynIII gene encoding a xylanase than existing heat-shock inducible promoter p (L), 1.2-fold and 3-fold production for xar gene encoding a arabinosidase than trc and T(7) promoter, respectively. The flow-in-heat technique created a rapid rise in temperature for effective induction of gene expression in bioreactor scale.

摘要

Hsh 基因表达系统旨在提高酶产量,降低大肠杆菌中基因表达诱导的成本。Hsh 系统的载体通过将合成的热激启动子与合成的终止子以及源自 pUC19 的复制起点结合而构建,其中外源基因的表达受大肠杆菌替代 sigma 因子 sigma(32)调控。相比之下,Hsh 启动子使编码木聚糖酶的 xynIII 基因的产量比现有热激诱导启动子 p(L)高 2.4 倍,使编码阿拉伯糖苷酶的 xar 基因的产量比 trc 和 T(7)启动子分别高 1.2 倍和 3 倍。流加热技术在生物反应器规模上快速升温,有效地诱导基因表达。

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