Renaturation and purification of ApxII toxin of Actinobacillus pleuropneumoniae

被引：3
作者：
Wang, Chunlai
Liu, Siguo
Peng, Yonggang
Shao, Meili
Wang, Yong
Gong, Qiang
Chang, Yuehong
Liu, Jiandong
Liu, Huifang
Liu, Di
Kong, Xiangang
机构：
[1] Chinese Acad Agr Sci, Vet Res Inst, Natl Key Lab Vet Biotechnol, Div Bacterial Dis, Harbin 150001, Peoples R China
[2] Heilongjiang Acad Agr Sci, Anim Husbandry Res Ctr, Harbin 150086, Peoples R China
[3] NE Forestry Univ, Harbin 150040, Peoples R China
来源：
PROTEIN EXPRESSION AND PURIFICATION | 2007年 / 52卷 / 02期
关键词：
expression; renaturation; purification; ApxII toxin; hemolytic activity;
D O I：
10.1016/j.pep.2006.10.026
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
ApxII toxin is the only Apx toxin that is produced by Actinobacillus pleuropneumoniae serotype 7. In order to determine whether the recombinant ApxII that derived from Escherichia coli (E. coli) expression is faithful to the natural ApxII so that can be used as additional component in vaccine preparation, the structure gene apxIIA of ApxII toxin was expressed in E coli with prokaryotic expression vector pGEX-6p-1 (formed pGEX-6p-A). pGZRS-C which is A. pleuropneunionicte-E. coli shuttle vector pGZRS-38 expressing the post-transcriptional activation gene apxII C was co-expressed with pGEX-6p-A. The expression product of rApxII A formed inclusion. The inclusion protein was oxidized, refolded and restored hemolytic activity after denaturation, renaturation and purification. The result indicated that E. coli expressed recombinant ApxII toxin has good fidelity, which makes it possible to produce this valuable antigen for vaccine preparation or diagnosis. (c) 2006 Elsevier Inc. All rights reserved.
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收藏
页码：441 / 445
页数：5
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