High yield expression of duck hepatitis A virus VP1 protein in Escherichia coli, and production and characterization of polyclonal antibody
文献类型: 外文期刊
第一作者: Li, Chuanfeng
作者: Li, Chuanfeng;Chen, Zongyan;Meng, Chunchun;Li, Lu;Liu, Guangqing
作者机构:
关键词: Duck hepatitis A virus;Codon optimization;Escherichia coli;VP1
期刊名称:JOURNAL OF VIROLOGICAL METHODS ( 影响因子:2.014; 五年影响因子:2.001 )
ISSN: 0166-0934
年卷期: 2013 年 191 卷 1 期
页码:
收录情况: SCI
摘要: VP1 protein, the capsid protein of duck hepatitis A virus (DHAV), contains critical epitopes for inducing a protective immune response. Due to its low-level expression in Escherichia coli (E. coli), the function of this protein is poorly characterized. In this study, a codon-optimized VP1 gene was chemically synthesized in terms of the codon usage bias in E. coli and subcloned into pET32a (+) to increase its expression. The recombinant VP1 fusion protein was purified from inclusion body by Ni2+ affinity chromatography His-Bind Resin and used to raise the rabbit anti-DHAV-VP1 polyclonal antibody. The expression of the codon-optimized VP1 gene in E. coli was significantly increased when compared to the wild-type VP1 gene, having an at least 17-fold increase. Western blot analysis showed that the recombinant protein was recognized by the rabbit anti-DHAV polyclonal antibody. Western blot also demonstrated that the rabbit anti-DHAV-VP1 polyclonal antibody could recognize the purified VP1 fusion protein specifically, and in the indirect immunofluorescent assays (IFA), the antibody was able to probe the VP1 protein in DHAV-1 infected cells. In conclusion, codon optimization increased dramatically DHAV VP1 expression in E. cob and the His-tagged VP1 fusion protein showed good antigenicity and immunogenicity. (C) 2013 Elsevier B.V. All rights reserved.
分类号:
- 相关文献
作者其他论文 更多>>
-
Proteomic analysis to explore potential mechanism underlying pseudomale sperm defect in Cynoglossus semilaevis
作者:Li, Xihong;Li, Lu;Cui, Zhongkai;Li, Ming;Sun, Xuexue;Li, Zhijie;Chen, Zhangfan;Ding, Lanqing;Xu, Wenteng;Li, Xihong;Cui, Zhongkai;Li, Ming;Chen, Zhangfan;Xu, Wenteng;Li, Lu;Xu, Dongdong;Xu, Wenteng
关键词:Cynoglossus semilaevis; Spermatogenesis; Proteomic profile; Chaperone; UPS
-
Discrimination of superficial lymph nodes using ultrasonography and tissue metabolomics coupled with machine learning
作者:Li, Lu;Wang, Xinyue;Deng, Hongyan;Lu, Wenjuan;Zhou, Yasu;Ye, Xinhua;Li, Yong;Wang, Jie
关键词:lymph nodes; lymphoma; metastasis; ultrasonography; metabolomics
-
MEHP induced mitochondrial damage by promoting ROS production in CIK cells, leading to apoptosis, autophagy, cell cycle arrest
作者:Li, Lu;Han, Bing;Yu, Yanbo;Lin, Hongjin;Li, Wan;Liu, Yufeng;Lin, Hongjin
关键词:MEHP; ROS; Mitochondrial damage; Apoptosis; Autophagy; Cell cycle arrest
-
Insight into the changes in structural, physicochemical, and probiotic properties of yam peel polysaccharides subjected to different physical-assisted extraction methods
作者:Lu, Jinghui;Li, Dingjin;Duan, Zhenhua;Lu, Jinghui;Yu, Yuanshan;Xu, Yujuan;Li, Lu;Cheng, Lina;Peng, Jian;Chang, Hong;Lu, Jinghui
关键词:Chinese yam peel polysaccharides; Physicochemical properties; Surface morphology; Functional activity
-
The interaction between soluble dietary fiber and bound phenolic from litchi pomace, and its effect on gut microbiota and inflammation
作者:Chen, Xiaowei;Xu, Hanting;Xu, Yujuan;Wu, Jijun;Zou, Bo;Li, Lu;Yu, Yuanshan;Hu, Tenggen
关键词:Litchi pomace soluble dietary fiber; Bound phenolic; Simulating digestion; Antioxidant potential; Anti-inflammatory potential
-
Optimizing Propylene/Propane Sieving Separation through Gate-Pressure Control within a Flexible Organic Framework
作者:Li, Lu;Xiang, Fahui;Li, Yunbin;Yang, Yisi;Yuan, Zhen;Chen, Yanting;Yuan, Furong;He, Lei;Xiang, Shengchang;Chen, Banglin;Zhang, Zhangjing;Xiang, Fahui
关键词:C3H6/C3H8 Sieving Separation; Gate pressure control; Hydrogen-Bonded Organic Framework; Temperature Dependent
-
The hypoglycemic effect of mulberry (Morus atropurpurea) fruit lacking fructose and glucose by regulation of the gut microbiota
作者:Wang, Ya-Ting;Wu, Ji-Jun;Yu, Yuan-Shan;Wen, Jing;Zou, Bo;Li, Lu;Peng, Jian;Cheng, Li-Na;Bu, Zhi-Bin;Xu, Yu-Juan;Hu, Teng-Gen;Wang, Ya-Ting;Wu, Hong;Yu, Yuan-Shan;Yu, Yuan-Shan;Zou, Bo;Xu, Yu-Juan;Hu, Teng-Gen
关键词: