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《Journal of Anhui Agricultural Sciences》 2009-10
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Expression of Porcine Reproductive and Respiratory Syndrome Virus ORF7 Gene and Purification and Immun ological Activity Analysis of the Recombinant Protein

ZHANG Yong-fu et al(College of Animal Science and Technology,Inner Mongolia University for the Nationalities,Tongliao,Inner Mongolia 028000)  
[Objective] The aim of this study was to realize efficient expression of the porcine reproductive and respiratory syndrome virus(PRRSV) ORF7 gene in genetic engineering bacteria and analyze the immun ological activity of the recombinant protein after purification.[Method]The constructed recombinant expression vector pET-ORF7 was transformed into Escherichia coli BL21(DE3) and induced by IPTG under the optimal condition.After analysis by SDS-PAGE and Western Blot,the expression products were purified by Ni-NTA His·Bind Resin chromatographic column under denaturing condition and renatured by gradient dialysis.Subsequently,the immun ological activity of the renatured recombinant protein was detected by Western Blot and indirect ELISA.[Result] The recombinant plasmid pET-ORF7 expressed in E.coli successfully,and the fusion protein was in the form of inclusion body.By SDS-PAGE detection,the molecular weight of the expression protein was approximate 33 kD,according with the expectation.Analysis by Bandscan software showed that the expressed fusion protein was about 50% of total bacterial protein of BL21(DE3).Western Blot and indirect ELISA detection showed that the renatured protein could react with PRRSV positive serum specifically,indicating its good immun ological activity.[Conclusion] This study lays a foundation for the preparation of PRRSV monoclonal antibody and diagnostic kit.
【Fund】: 北京市农林科学院青年基金项目(2007020122);; 北京市科委猪高致病性蓝耳病防控技术子课题
【CateGory Index】: S858.28
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【Citations】
Chinese Journal Full-text Database 2 Hits
1 LU Wei-wei1,JIANG Ping1,TANG Tai-shan2,LI Yu-feng1,ZHANG Chang-yin2(1.Key Laboratory of Animal Disease Diagnostics and Immunology,Ministry of Agriculture,Nanjing Agricultural University,Nanjing,Jiangsu,210095,China;2.Jiangsu Entry-Exit Inspection and Quarantine Bureau,Nanjing,Jiangsu,210001,China);Genetic Variance Analysis of ORF7 Gene Sequences of PRRSV[J];Progress in Veterinary Medicine;2007-11
2 ZHANG Yongfu1,2,3,MA Guowen1,LIU Yuehuan2,LIU Yonghong2,4,LI Minggang3, ZHANG Qiuyu3,LIU Feng3 (1.College of Animal Science and Technology,Inner Mongolia University For the Nationalities,Tongliao Inner Mongolia 028000;2. Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100089;3.Beijing DIHO Biomedicai Science &Technology Co.,Ltd, Beijing 100043;4. College of Animal Science and Medicine, Inner Mongolia Agriculture University,Huhhot Inner Mongolia 010018 );Construction and Sequence analysis of the Prokaryotic Expression Vector for ORF7 Gene of Porcine Reproductive and Respiratory Syndrome Virus[J];Chinese Journal of Animal Health Inspection;2008-12
【Co-citations】
Chinese Journal Full-text Database 2 Hits
1 YAO Xue-jing1,ZHAN Song-he2,ZHU Liang-qiang2,QI Ke-zong1,WANG Fei2,WANG Ai-ping1 (1.College of Animal Science and Technology,Anhui Agricultural Univresity,Hefei,Anhui,230036,China;2.Anhui Centre for Animal Diseases Control and Prevention,Hefei,Anhui,230022,China);Cloning and Sequence Analyses of the NSP2 and ORF7 Genes of PRRSV Strains in Anhui Province[J];Progress in Veterinary Medicine;2010-05
2 JIAN Zi-jian1,SHENG Zhuo-jun2,SU Gui-cheng1,MA Su-zhen1,WANG Xiao-ping1,HU Jun2,WANG Wei2,MA Xiao-yan2,RAN Duo-ling1(College of Animal Medicine,Xinjiang Agricultural University,83005,China;2.Xinjiang Institute of Animal health Supervision,Urumqi 830063,China);Identification and Isolation of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus in Xinjiang[J];Xinjiang Agricultural Sciences;2008-06
【Secondary Citations】
Chinese Journal Full-text Database 3 Hits
1 PENG Chang-ling, GAO Song, LIU Xiu-fan(Key Laboratory of Animal Infectious Disease, Ministry of Agriculture/Yangzhou University, Yangzhou 225009,China);Development of indirect immunofluorescent assay to detect porcine reproductive and respiratory syndrome virus[J];Chinese Journal of Veterinary Science and Technology;2005-04
2 Guo Baoqing(Harbin Veterinary Research Institute,the Chinese Academy of Agricultural Sciences);Isolation and Identification of Porcine Reproductory and Respiratory Syndrome(PRRS) Virus from aborted fetuses suspected of PRRS[J];;1996-02
3 CUI Shang-jin,JIANG Jian-hong,ZHOU Yan-jun,QIU Hua-ji,TONG Guang-zhi(Harbin Veterinary Research Institute of CAAS 150001,China);The development and preliminary application of immuno-gold filtration assay for porcine reproductive and respiratory syndrome[J];Chinese Journal of Preventive Veterinary Medicine;2005-03
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