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《Progress in Veterinary Medicine》 2010-01
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Expression and Antigenicity Analysis of the Omp31 Protein of Brucella melitensis

XU Jie1,2,WANG Yu-fei2,WANG Zhou-jia2,QIAO Feng2,ZHONG Zhi-jun 2,3,DU Xin-ying2,ZHAO Jin2,QU Qing2,GAO Lan1,CHEN Ze-liang2(1.College of Life Science,Lanzhou University,Lanzhou,Gansu,730000,China;2.Institute of Disease Control and Prevention,Academy of Military Medical Science,Beijing,100071,China;3.College of Veterinary Medicine,Nanjing Agricultural University,Nanjing,Jiangsu,210095,China)  
In order to clone and express the Omp31 protein of B.melitensis in E.coli,purify the expressed protein and detect its immunogenicity.A gene recoding outer membrane protein 31-34 ku(Omp31) was amplified from the genomic DNA of B.melitensis by PCR.The amplified fragments were digested with BamHⅠ and SalⅠ,and then cloned to the vector pET32a.The constructed recombinant plasmid pET32a-Omp31 was transformed to E.coli ER2566(DE3) and was induced to express the fusion protein.Then the protein was purified by histidine-binding resin column chromatography,and the immunogenicity was detected by Western blot.The PCR product of Omp31 gene was cloned to the vector pET32a,and the recombinant vector was confirmed by colony PCR identification,recombinant vector digested identification and sequencing analysis.It was successfully expressed in E.coli ER2566(DE3) as a fusion protein with histidine at the presence of IPTG,and a specific protein band of 43 ku was found when identified by SDS-PAGE.Western blot showed good immunoreactivity of the expressed product.Omp31 was successfully cloned and expressed,and the purified fusion protein had immunogenicity.This study provides a solid foundation for the further study on the function of proteins and brucellosis diagnostic antigens.
【Fund】: 国家自然科学基金项目(30600024);; 国家"863"资助项目(2007AA02Z412)
【CateGory Index】: S852.614
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【Citations】
Chinese Journal Full-text Database 1 Hits
1 Yufei Wang**, Zeliang Chen**, Feng Qiao, Xinying Du, Leili Jia, Jing Yuan, Liuyu Huang* (Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071);Recombinational Cloning of Large DNA Fragments of Bacterial Chromosome by Plasmid Rescue[J];Acta Microbiologica Sinica;2008-04
【Secondary Citations】
Chinese Journal Full-text Database 1 Hits
1 WANG Yu-Fei CHEN Ze-Liang ZHAO Hong-Qing YUAN Xi-Tong HUANG Liu-Yu ZHANG Ling LIU Jing-Mei SONG Hong-Bin(Institute of Disease Control and Prevention, Academy of Military Medical Sciences,Beijing 100071);Construction of Brucella Unmarked Deletion Mutant by Using Conventional Cloning Vector as Suicide Plasmid[J];Microbiology;2007-04
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