Cloning of E Gene of PRRS Virus and Construction of Its Expressing Vectors

doi:10.3321/j.issn:1000-7091.2006.01.007

Abstract

Abstract: The E gene fragments of PRRS strain BJ-4 have been amplified and cloned into plasmid pBV221 and pPIC9K.Recombinant plasmid was constructed and transformed into the JM109.The result indicated that the recombinant vectors were successfully constructed.Positive clones of target genes were screened by PCR and identified with restriction endo-nuclease analysis.The positive clone of recombinant prokaryotic expression vector was induced and expressed with ELISA showed that it could react with the antibody to PRRSV.The results showed that E Protein of PRRSV was expressed in prokaryote system.The prokaryotic expression vector and eucaryotic expression vector were successfully constructed,which would be useful for production of gene vaccination in future.

Key words: PRRSV, Gene cloning, Prokaryotic expression, Eucaryotic expression

CLC Number:

Q342

YUAN Bo-hua, JIANG Ji-zhi, LIU Hong-mei. Cloning of E Gene of PRRS Virus and Construction of Its Expressing Vectors[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2006, 21(1): 27-30. doi: 10.3321/j.issn:1000-7091.2006.01.007.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.3321/j.issn:1000-7091.2006.01.007

http://www.hbnxb.net/EN/Y2006/V21/I1/27

References

[1] Wensvoort G,Tepstra C,Pol J M A,et al.Mystery swine disease in the Netherlands:the isolation of Lelystad virus [J].Veterinary Quarterly,1991,13:121-130.
[2] Terpstra C,Wensvoort G,Pol J M A.Experimental reproduction of porcine epidemic abortion and respiratory syndrome(mystery swine disease)by infection with Lelystad virus [J].Vet Q,1991,13:131-136.
[3] Mengeling W L,Lager K M,Vorwald A C.Clinical effects of porcine reproductive and respiratory syndrome virus on pigs during the early postnatal interval[J].Am J Vet Res,1998,59(1):52-55.
[4] 郭宝清,陈章水,刘文兴,等.从疑似PRRS流产胎儿分离PRRSV的研究[J].中国畜禽传染病,1996,18(2):1-4.
[5] 乔传玲,郑厚旌,甘孟侯,等.反转录聚合酶反应检测猪繁殖与呼吸综合征病毒的研究[J].畜牧兽医学报,2000,31(3):262-266.
[6] 杨汉春,管山红,尹晓敏,等.猪繁殖与呼吸综合征病毒的分离与鉴定[J].中国兽医杂志,1997,23(11):3-5.
[7] Pringle C R.Virus Taxonomy 1997-Proceedings of the Excutive Committee of the ICTV [J].ArchVirol,1997,142 (8):1727-1733.
[8] Meulenberg J J M,Besten A P,de Kluyver E P,et al.Characterization of proteins encoded by ORFs 2 to 7 of Lelystad virus [J].Virology,1995,206:155-163
[9] Sur J H,Doster A R,Osorio F A.Apoptosis induced in vivo during acute infection by porcine reproductive and respiratory syndrome virus[J].Vet Pathol,1998,35(6):506-514.
[10] Delputte P L,Vanderheijden N,Nauwynck H J,et al.Involvement of the matrix protein in attachment of porcine reproductive and respiratory syndrome virus to a heparinlike receptor on porcine alveolar macrophaGes [J].J Virol,2002,76(9):4312-4320.
[11] 奥斯伯F,布伦特R,金斯敦R E.精编分子生物学实验指南[M].颜子颖等,译(第3版).北京:科学出版社,1999.
[12] Sambrook J,Russell D W.Moecular cloning:A Laboratory Manual 3 rd ed[M].New York:Cold Spring Harbor Laboratory,2001.8:40-55.

[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]	HOU Ruize, HOU Yuxiang, XU Xiaoyong, LI Xuan, LI Meilan. Cloning and Expression Analysis of CYP Homologous Genes in Brassica rapa ssp. chinensis [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(5): 45-51.
[3]	JIN Yifeng, GAO Yansong, WANG Qi, WANG Mengmeng, ZHAO Di, XIONG Yi, CHEN Yang. *Cloning of SnRK2.4* Gene and It's Response to Abiotic Stress in Kentucky Bluegrass** [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(5): 9-15.
[4]	ZHANG Dongmei, FENG Yayan, XIU Zhijun, YANG Chunfang, DU Meie, LI Dezhou, ZHANG Xiaoyu. Cloning and Bioinformatics Analysis of Resistant Gene StWRKY11 Induced by Sodium Silicate in Potato by Rhizoctonia solani [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(5): 194-203.
[5]	ZHENG Yusi, WANG Pei, GUO Ying, BAI Lirong, YU Dahui, ZHAO Sen. Molecular Cloning and Expression Analysis of PfCLEC17A in Pearl Oyster Pinctada fucata [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(5): 230-238.
[6]	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.
[7]	JIA Zhiqiang, XU Yunyu, GAO Xue, TAO Hongzheng, CHEN Zengmin, LIU Yating, LI Yongzhong. *Cloning,Subcellular Positioning of Pepper CaWRKY30* Transcription Factor, and Its Expression Analysis by Tomato spotted wilt orthotospovirus Infected** [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(3): 186-192.
[8]	ZHAO Huiying, YANG Guangkai, GAO Yan, ZHANG Xiaojun, HAO Yanyan. *Cloning of Gene MdSGR2* from Apple and Construction of Plant Overexpression Vector** [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(2): 42-48.
[9]	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.
[10]	JIA Weizhe, JIAO Bo, WANG Jiao, CHEN Wenye, YANG Fan, LIU Yongwei, DONG Fushuang, ZHAO Liqun, ZHOU Shuo. *Cloning and Expression Analysis of Calmodulin-like Protein TaCML8-A* Gene in Wheat** [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(1): 1-8.
[11]	TANG Wenfang, XU Shengsheng, DUAN Yanbi, LONG Wenhong, YANG Rongping, ZHANG Xuemei, MENG Jingui. Cloning of DoIPTs Genes in Yam and Its Expression Analysis During Bulbil Germination [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(1): 27-34.
[12]	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.
[13]	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.
[14]	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.
[15]	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.

Please choose a citation manager

Content to export