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標題: 共表現TorD對利用雙精胺酸轉位系統轉位基因重組蛋白之影響
The effect of TorD coexpression on the translocation of recombinant protein via the TAT pathway
作者: 李思禹
Li, Si-Yu
關鍵字: twin-arginine translocation coexpression TorD GFP;雙精胺酸 轉位 共表現 綠色螢光蛋白 TorD
出版社: 化學工程學系
Twin-arginine translocation pathway is capable of secreting fully folded proteins into the periplasm of Gram-negative bacteria and may thus be an ideal system for the expression of active cofactor-containing proteins. However, the applications of Tat system for such purpose have been plagued by low translocation efficiencies. In this study, we demonstrate that the coexpression of a soluble chaperone, TorD, in conjunction with the TorA signal peptide, the translocation efficiency of GFP can be enhanced by more than three-fold. The enhancement in translocation efficiency is believed to be a result of reduced proteolysis mediated by the binding of TorD toward the TorA signal peptide. We believe this approach can be further exploited for the expression and secretion of other heterologous proteins as well as traditional Tat substrate proteins.

雙精胺酸轉位系統(Twin-arginine translocation, Tat)為一新穎之蛋白輸送系統,其特點在於可選擇性地將摺疊完成之蛋白自細胞質區輸送至細胞間質區,因此簡化後續蛋白回收的程序。然而,過去的文獻指出,利用大腸桿菌雙精胺酸轉位系統輸送基因重組蛋白質的效率往往不甚理想。本研究利用共表現TorD蛋白增進利用雙精胺酸轉位系統輸送TGS的效率。
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