从大肠杆菌中分泌外源基因产物至培养基：利用ompF基因分泌人β-内啡肽。
Secretion into the culture medium of a foreign gene product from Escherichia coli: use of the ompF gene for secretion of human beta-endorphin.
作者信息
Nagahari K, Kanaya S, Munakata K, Aoyagi Y, Mizushima S
出版信息
EMBO J. 1985 Dec 16;4(13A):3589-92. doi: 10.1002/j.1460-2075.1985.tb04121.x.
Abstract
The ompF gene codes for a major outer membrane protein of Escherichia coli. A plasmid was constructed in which the structural gene for human beta-endorphin is preceded by the upstream region of the ompF gene consisting of the promoter region and the coding regions for the signal peptide and the N terminus of the OmpF protein. When the plasmid was introduced into E. coli N99, and OmpF-beta-endorphin fused peptide was synthesized and secreted into the culture medium through both the cytoplasmic and outer membranes. The OmpF signal peptide was cleaved correctly during the secretion, indicating that the export of the fused protein across the cytoplasmic membrane was dependent on the signal peptide. The secretion into the culture medium was apparently selective. Neither beta-lactamase nor alkaline phosphatase (both are periplasmic proteins) appeared in the culture medium in significant amounts. The mode of passage of the fused peptide across the outer membrane is discussed.
摘要
ompF基因编码大肠杆菌的一种主要外膜蛋白。构建了一种质粒，其中人β-内啡肽的结构基因之前是ompF基因的上游区域，该区域由启动子区域以及OmpF蛋白信号肽和N端的编码区域组成。当将该质粒导入大肠杆菌N99时，合成了OmpF-β-内啡肽融合肽，并通过细胞质膜和外膜分泌到培养基中。在分泌过程中，OmpF信号肽被正确切割，这表明融合蛋白穿过细胞质膜的输出依赖于信号肽。分泌到培养基中的过程显然具有选择性。β-内酰胺酶和碱性磷酸酶（两者均为周质蛋白）均未大量出现在培养基中。文中讨论了融合肽穿过外膜的方式。
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本文引用的文献
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