Prokaryotic Expression of GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus Hn-1/06 Strain

doi:10.7668/hbnxb.2009.01.023

Abstract

Abstract: One pair of specific primers was designed according to the sequence of PRRSV ORF3 gene published inCnBank(Accession number PRU87392),and was used to ampl the ORF3 gene from a PRRSV Hn 1/ 06 strain by RT-PCR method,resulting in a 765 gene fragment. The RT-PCR product was directly cloned into the pTG19-T vector andthe recombinant plasmid was identified by enzyme digesting and DNA sequencing. The positive clone was designated asp TG19- T-GP3.After being cloned into the prokaryotic expression vector pET-32a,the ORF3 gene was successfully ob-twined by the induction of IPTG. Western blot test confirmed that the expressed fusion protein could specifically react withthe antiserum against PRRSV. It could be used for the further investigation on immunogenicity and function of GP3 of PRRSV.

Key words: PRRSV, GP3, Prokaryotic expression

CLC Number:

S852.65

WANG Zhong-ming, LI Su-ping, XIA Ping-an, YIN Yan-tao, LI Wei-juan, QIU Huang, LIU Ming-li, LIU Yu-song, CUI Bao-an. Prokaryotic Expression of GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus Hn-1/06 Strain[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(1): 103-107. doi: 10.7668/hbnxb.2009.01.023.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2009.01.023

http://www.hbnxb.net/EN/Y2009/V24/I1/103

References

1 Hall W V. R}rcine reproductive and respiratory syndrome (PRRS) virus a significant disease of pigs [J].Aust Vet J， 2005,83 (5)：260一261.
2 Van Nieuw stadt A P,Meulenberg J M,Van Essem 7andberge- nA, et al. Proteins encoded by open reading frames 3 and 4 of the genome of Lelystad virus (Arteriviridae) are structural pro- teins of the virion [J]. J Virol,1996,70：4767一4772.
4 Meulenberg J M, Petersen-den Besten A,De HIuyver E P，et al. Characterization of proteins encoded by ORFs 2 to 7 of the lystad virus [J].Virology,1995,206：155一163.
4 Plana duran J,Climent I,Sarraseca J，et al. Baculovrius ex- pression of proteins of porcine reproductive and respiratory syndrome virus strain Olot/ 91 involvement of ORF3 andORFS proteins in protection[J].Virus Cenes,1997,14:19-29.
5 杨金雨，仇华吉，童光志，等.猪繁殖与呼吸综合征疫苗研究进展[J].动物医学进展，2005,26 (5)：28一31.
6 梁国栋.最新分子生物学实验技术[M].北京:科学出版社，2001：30一43.
7 萨姆布鲁克J,D. W.拉塞尔.分子克隆实验指南[M].黄培堂等译.第3版.北京:科学出版社，2002：1256 -1259.
8 萨姆布鲁克J沸里奇E F,曼尼阿蒂斯T.分子克隆实验指南[M]金冬雁等译.第2版.北京:科学出版社，1998：16一55,880一887.
9 秦睿玲，张西臣，李建华，等.柔嫩艾美球虫问接FL 1SA检测方法的建立[J]中国兽医科技，2004,34 (7)：60 -63.
10 胡慧.猪瘟病毒E2基因主要抗原区的表达及其表达产物的问接FI,1S、诊断方法研究[D].杨凌:西北农林科技大学，2004.
11 Murtaugh M P, F1am M R，Kakach L T. Comparison of the structural protein coding sequences of the VR22332 andLelystad virus strains of the PRRS virus [J].Arch Virol， 1995,140：1451一1460.
12 Dea S，Cegnon C A,Mardassi H，et al. Current knowle on the structural proteins of porcine reproductive and respi- ratory syndrome(PRRS) virus:comparison of the North American and European isolates [J].Arch Viro1,2000,145 (4)：659一688.
13 Meulenberg J M,PetersemDen Besten A. Identification and characterization of a sixth structural protein of Lelystad virus ahe glycoprotein C2 encoded by ORF2 is incorporated in virus particle [J].Virology,1996,225：44一51.
14 Wieczorel Krohmer M, Weiland F, Conzelinann K, et al. Rrcine reproductive and respiratory syndrome virus (PRRSV):rmnoclonal antibodies detect comrmn epitopes on two viral proteins of European and U. S. isolates[J].Vet- 1VTi-crobiol,1996,51(3一4)：257一266.
15 Mardassi H, Ornin P, Qgmn C A, et al. A subset of porcine reproductive and respiratory syndrome virus GP3 glycoprotein is released into the culture medium of cells as a non-virion associated and membrane free(soluble) form [J].J Virol， 1998,72：6298一6306.

[1]	XIAO Shikui, LI Fang, ZHANG Wenting, LÜ Shufang, SHI Guoan, WU Jiang, FAN Bingyou. *Cloning and Prokaryotic Expression of ACS* Gene from Paeonia lactiflora Pall.** [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(5): 36-44.
[2]	WANG Ya'nan, LIANG Yanyan, YAN Wenpei, ZHANG Yinping, LI Qiang, WU Xiuju. Cloning and Prokaryotic Expression Analysis of AhOMT1 Gene from Asarum heterotropoides [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(4): 62-70.
[3]	LIAN Kaiqi, JI Shuang, ZHOU Lingling, SHI Zhiqi, WANG Fei, ZHANG Yuanchen. Cloning and Prokaryotic Expression of a clip-type Serine Protease Gene from Mythimna separate [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(1): 195-201.
[4]	FAN Huifen, SUN Tianjie, SU Weihua, XIAO Fuming, ZHANG Jie, WANG Dongmei. Preparation of GmWRKY50 Protein Antibody and Analysis of Its Protein Level Expression [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(6): 28-34.
[5]	XU Huan, DUAN Shengwen, FENG Xiangyuan, ZHENG Ke, YANG Qi, WANG Qiming, CHENG Lifeng, PENG Yuande. High Level Expression of Pectate Lyase Gene from Microbiology for Ramie Degumming and Its Bioinformatics Analysis [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(5): 18-23.
[6]	LEI Qidong, SUN Xudong, XU Huini. *Cloning and Prokaryotic Expression of Arabidopsis AtTCP4* Gene** [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(5): 24-28.
[7]	JIANG Nan, ZHENG Qianqian, LI Qianwen, TU Jian, SONG Xiangjun, SHAO Ying, LIU Hongmei, QI Kezong. Prokaryotic Expression Vector Construction,Expression,Purification and Identification of the APEC Virulence Gene ygeG [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(5): 184-190.
[8]	YU Tianfei, XIE Pengyu, SUN Wanshu, LI Jing, YIN Haichang, LI Ming, YU Zhidan. *Prokaryotic Expression of Variable Domain of Monoclonal Antibody Against Goose parvovirus* NS1 Protein** [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(5): 211-215.
[9]	WANG Haibo, LI Furong, YANG Jincui, GAO Yong, GUO Junyun. Cloning and Prokaryotic Expression of Phosphoglucomutase Genes cPGM and pPGM from Jatropha curcas [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 23-30.
[10]	LI Moxiao, BIAN Zhe, ZHOU Qihui, LIU Yuwei, GONG Xiaodong, GU Shouqin, HAN Jianmin. Analysis of the Gene Structure,Prokaryotic Expression and Expression Pattern of StRTG2 in Setosphaeria turcica [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 191-196.
[11]	LIU Qing, SUN Tingzhao, GAI Shupeng, ZHANG Yuxi, LIU Chunying. Characteristic and Expression Analysis of a Transcription Factor PsZFP1 in Tree Peony [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(3): 67-73.
[12]	LI Jiahao, XIE Minqiu, WAN Chuanyin, ZUO Ruijie, LU Liming, LI Liqin. *Prokaryotic Expression of Transcription Factor StTCP13* in Potato and Its Function of Salt Stress** [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(2): 33-39.
[13]	HU Yueqing, WANG Ruozhong, HUANG Zhigang, XIAO Langtao. *Cloning and Prokaryotic Expression of Transcription Factor StWRKY57* Gene in Potato** [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(1): 10-17.
[14]	LIU Dong, LAN Yanping, ZHANG Li, ZHAO Yan, QU Yunfang, HUANG Jinling. *Optimization of Prokaryotic Expression Conditions of Cotton GhPDP* Gene** [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(5): 55-61.
[15]	LUAN Qingdong, SUN Ju, LIU Junhua, ZHANG Benqing, YIN Yanbo, WANG Jianlin. *Cross Neutralization of Fiber Protein Against Four Kinds of Serotype fowl adenovirus* Prevalent in China** [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(5): 214-219.

Please choose a citation manager

Content to export