Modification with a phosphorylation tag of PKA in the TraT-based display vector of Escherichia coli

被引:14
作者
Chang, HH [1 ]
Lo, SJ [1 ]
机构
[1] Natl Yang Ming Univ, Sch Life Sci, Inst Microbiol & Immunol, Taipei 11221, Taiwan
关键词
bacterial display; TraT lipoprotein; rhodostomin; protein kinase A; thrombin;
D O I
10.1016/S0168-1656(99)00227-8
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We have previously developed the TraT display system to express the preS1 peptide of human hepatitis B virus (HBV) and the snake venom rhodostomin (RHO) on the surface of Escherichia coli. In this study, we modified the pT2 vector by adding a thrombin cutting site and a phosphorylation tag of protein kinase A before the multiple restriction enzyme sites. The modified vector allowed us to label the TraT fusion protein (TraT-RHO) with [P-32] and to increase the detection sensitivity of TraT-RHO expression bacteria binding to and being internalized into BHK-21 cells. After the thrombin cleavage, the isotope labeled RHO could be detected in a free form. We therefore suggest that the new version of pT2 vector, pT2-KL, will facilitate to identify the counterpart of displayed peptide. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:115 / 122
页数:8
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