Cell surface display of salmobin, a thrombin-like enzyme from Agkistrodon halys venom on Escherichia coli using ice nucleation protein
Cell surface display of salmobin, a thrombin-like enzyme from Agkistrodon halys venom on Escherichia coli using ice nucleation protein
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DOI：
10.1016/s0141-0229(00)00315-x
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发表时间：
2001-02-01
影响因子：
3.4
通讯作者：
Kim, EJ
中科院分区：
文献类型：
--
作者：
Jeong, HS;Yoo, SK;Kim, EJ
利用冰核蛋白在大肠杆菌上进行细胞表面展示，以开发一种新的重组真核蛋白表达系统。从蛇毒中提取的类凝血酶Salmobin被展示在与冰核蛋白(INP)C端融合的大肠杆菌表面。紫丁香假单胞菌的一种外膜蛋白。凝血酶裂解位点位于沙莫酶和INP之间。经SDS-PAGE、Western blotting、全细胞ELISA法和免疫荧光显微镜检测，证实细菌细胞表面存在沙莫宾。凝血酶切割后，对重组沙莫酶的类凝血酶活性进行了检测和验证。我们的结论是，基于INP的细胞表面展示可以作为一种新的真核蛋白表达系统。(C)2001 Elsevier Science Inc.保留所有权利。
Cell surface display on Escherichia coli using ice nucleation protein was performed in order to develop a new expression system for recombinant eukaryotic proteins. Salmobin, the thrombin-like enzyme obtained from Korean snake (Agkistrodon halys) venom was displayed on the surface of Escherichia coli fused to the C-terminus of the ice nucleation protein (INP). an outer membrane protein of Pseudomonas syringae. The thrombin cleavage site was inserted between salmobin and INP. The presence of salmobin on the bacterial cell surface was verified by SDS-PAGE, Western blotting, whole cell ELISA, and immunofluorescence microscopy. After thrombin cleavage the thrombin-like enzyme activity of recombinant salmobin was tested and verified. We concluded that INP-based cell surface display can be used as a novel expression system for eukaryotic proteins. (C) 2001 Elsevier Science Inc. All rights reserved.