Prokaryotic expression and polyclonal antibody preparation of σC protein of novel duck reovirus

Prokaryotic expression and polyclonal antibody preparation of σC protein of novel duck reovirus

1 College of Animal Science and Technology， Anhui Agricultural University， Hefei 230036， China;2 Shanghai Veterinary Research Institute， CAAS， Shanghai 200241， China

Online:2014-07-25 Published:2014-08-06

Abstract

Abstract: To investigate the immunogenicity of σC protein of a novel duck reovirus (NDRV TH11)， the σC gene was cloned into pET30a(+) and expressed in E.coli BL21(DE3). SDSPAGE and Western blot analysis showed that the expressed had good immunological activity. The polyclonal antibody was obtained from the rabbit immunized with purified recombinant protein and its titer was about 1∶20 000 by detection of indirect ELISA. Indirect immunofluorescence assay also demonstrated that the polyclonal antibody reacted specially with the native NDRV σC protein. This study will be helpful for preparation of peptide vaccines and σC function study.

Key words: novel duck reovirus, σC gene, prokaryotic expression, polyclonal antibody

BI Zhuangli1，2， ZHU Yingqi1，2， CHEN Zongyan2， LI Chuanfeng2， MENG Chunchun2， WANG Guijun1， LIU Guangqing2，*. Prokaryotic expression and polyclonal antibody preparation of σC protein of novel duck reovirus[J]. .

[1]	ZHAO Xiuping, WANG Shuang, YAN Xingyi, DUAN Qiang, ZHANG Shuai, CHEN Yongsheng, LI Guorui. Expression, purification and bioinformatics analysis of Magnaporthe oryzae MGG-01005 [J]. Acta Agriculturae Zhejiangensis, 2021, 33(3): 470-478.
[2]	SU Xuesi, ZHANG Yubao, WANG Ruoyu, WANG Yajun, TANG Guoliang, JIN Weijie. Prokaryotic expression of Plantago asiatica mosaic virus capsid protein and preparation of its polyclonal antibody [J]. Acta Agriculturae Zhejiangensis, 2021, 33(1): 104-111.
[3]	LIU Jinyu, HUANG Ying. Prokaryotic expression and antibody preparation of full-length and N-terminal half of SpTrz2 protein in Schizosaccharomyces pombe [J]. Acta Agriculturae Zhejiangensis, 2021, 33(1): 34-42.
[4]	HUANG Xiaozhen, QIAO Zhongquan, ZENG Huijie, LI Yongxin, HE Gang, WANG Xiaoming. Isolation and expression of floral organ development regulating gene LiFUL1 in Lagerstroemia indica L. [J]. , 2020, 32(7): 1206-1214.
[5]	CHEN Yunlu, SHAN Ying, LUO Hao, XU Jidong, ZHAO Lingyan, FANG Weihuan, LI Xiaoliang. Prokaryotic expression of porcine type Ⅲ interferon and studying on its antiviral activity [J]. , 2020, 32(5): 779-788.
[6]	PU Lusha, SU Shibo, CHEN Xiaohan, ZHAO Lili, CHEN Hongyan. Prokaryotic expression and phylogenetic analysis of ORF2 gene of goose astrovirus [J]. , 2020, 32(5): 789-797.
[7]	YUN Tao, HUA Jionggang, YE Weicheng, NI Zheng, CHEN Liu, ZHANG Cun. Development and application of a one-step real-time TaqMan-MGB RT-PCR assay for detection of novel duck reovirus [J]. , 2020, 32(4): 571-576.
[8]	WANG Yuanhong, XING Xue, LI Chuanfeng, ZHU Jie, WANG Yong, LIU Guangqing. Bioinformatics analysis and prokaryotic expression of FIPV AH1905 strain N gene [J]. , 2020, 32(3): 406-414.
[9]	WANG Xiaopeng, ZHAO Liang, LIU Zimin, BAI Caixia, YANG Kankan, ZHANG Da, SUN Pei, JIANG Shudong, LI Yongdong, WANG Yong. Prokaryotic expression and bioinformatics analysis of PPV7 Cap gene [J]. , 2020, 32(2): 200-209.
[10]	LI Xianchun, LU Yan, MAO Yaofang, YANG Haifeng, YU Haishan, MA Yonghua1, WAN Xuerui. Construction of prokaryotic expression vector of chicken Prnp gene and expression in Escherichia coli [J]. Acta Agriculturae Zhejiangensis, 2020, 32(12): 2138-2146.
[11]	ZHANG Haojie, LIU Mei, LI Chunyan, HE Ran, LAN Jingchao, LUO Li, GU Xiaobin, XIE Yue, YANG Guangyou. Prokaryotic expression and immunofluorescence localization of nucleoside diphosphate kinase gene from Dirofilaria immitis [J]. , 2019, 31(9): 1453-1460.
[12]	PAN Chuanyan, LIN Yong, FENG Pengfei, ZHANG Yongde, LUO Honglin. Prokaryotic expression and polyclonal antibody preparation of Hsc70 in Nile tilapia [J]. , 2019, 31(8): 1272-1279.
[13]	YUN Tao, HUA Jionggang, YE Weicheng, NI Zheng, CHEN Liu, ZHANG Cun. Proliferative characteristics of novel duck reovirus strain JDM10 in DF-1 cells [J]. , 2019, 31(5): 716-721.
[14]	LIU Zhengkui, WU Yuan, CHEN Lin, WANG Lei, MU Hongye, ZHU Xuhang, WANG Xiaodu. Prokaryotic expression and bioinformatics analysis of Nsp5 gene of porcine epidemic diarrhea virus [J]. , 2019, 31(4): 532-538.
[15]	PENG Qiqi, YANG Jian, LIAO Qiansheng, ZHANG Hengmu. Preparation and application of multiclonal antibody against plant cysteine proteinase [J]. , 2018, 30(6): 881-885.

Prokaryotic expression and polyclonal antibody preparation of σC protein of novel duck reovirus

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments