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《Chinese Journal of Cellular and Molecular Immunology》 2007-01
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Construction and prokaryotic expression of recombinant gene EGFRvIII-HBcAg and immunogenicity analysis of the fusion protein

DUAN Xiao-yi~1, WANG Jian-sheng~1, GUO You-min~ 2* , HAN Jun-li~2, WANG Quan-ying~3, YANG Guang-xiao~3 ~1Department of Nuclear Medicine, the First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061; ~2Department of Image Center, the Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710004, China  
AIM: To construct recombinant prokaryotic expression plasmid pET28a(+)/c-PEP-3-c and evaluate the immunogenicity of the fusion protein. METHODS: cDNA fragment encoding PEP-3 was obtained from pGEM-T Easy/PEP-3 and inserted into recombinant plasmid pGEMEX/ HBcAg . Then it was subcloned in prokaryotic expression vector and transformed into E.coli BL21(DE3). The fusion protein was expressed by inducing IPTG and purified by Ni~ 2+ -NTA affinity chromatography. BALB/c mice were immunized with fusion protein and the antibody titre was determined by indirect ELISA. RESULTS: The recombinant gene was confirmed to be correct by restriction enzyme digestion and DNA sequencing. After prokaryotic expression, fusion protein existed in sediment and accounted for 56% of all bacterial lysate. The purified product accounted for 92% of all protein and its concentration was 8 g/L. The antibody titre in blood serum reached 1∶16000 after the fourth immunization and reached 1∶2.56×10~5 after the sixth immunization. The titre of anti-PEP-3 antibody reached 1∶1.28×10~5 and the titre of anti-HBcAg antibody was less than 1∶4×10~3. CONCLUSION: Fusion gene PEP-3-HBcAg is highly expressed in E.coli BL21. The expressed fusion protein can induce neutralizing antibody with high titer and specificity, which lays a foundation for the study of genetically engineering vaccine for malignant tumors with the high expression of EGFRvIII.
【Fund】: 国家自然科学基金资助项目(30600744)
【CateGory Index】: R730.5
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【References】
Chinese Journal Full-text Database 1 Hits
1 LIU Bo;SUN Ze-qiang;LIU Qing-yong;CHEN Jie;WANG Si-jun;YANG Guang-xiao;WANG Quan-ying;School of Medicine of Shandong University;Department of Urinary Surgery,Ji'nan Central Hospital Affiliated to Shandong University;Xi'an Huaguang Biological Engineering Co.Ltd.;;Construction and prokaryotic expression of recombinant gene G250 antigenic peptide-HBcAg and the immunogenicity analysis of the fusion protein[J];西安交通大学学报(医学版);2014-01
【Citations】
Chinese Journal Full-text Database 2 Hits
1 Duan Xiaoyi~1,Wang Jiansheng~2,Guo Youmin~3,Liu Min~3,Zhou Bin~3,Wang Quanying~4,Yang Guangxiao~4(1.Department of Nuclear Medicine;2.Department of Oncosurgery,First Affiliated Hospital,Medical School of Xi'an Jiaotong University,Xi'an 710061;3.Image Centre,SecondAffiliated Hospital,Medical School of Xi'an Jiaotong University,Xi'an 710004;4.Xi'an Huaguang Biological Engineering Company,Xi'an 710025,China);Clone of cDNA encoding PEP-3 epitope of epidermal growth factor receptor type Ⅲ mutant[J];西安交通大学学报(医学版);2006-02
2 FENG Gai-feng~1, HU Hai-tao~1, LI Yue-ying~1, HAN Hua~1, WANG Quan-ying~2, YANG Guang-xiao~2~1Department of Human Anatomy, Medical College, Xi'an Jiaotong University, Xi'an 710061; ~2Xi'an Huaguang Biotechnology Company, Xi'an 710025, China;Construction and prokaryotic expression of recombinant gene c-Aβ-c and the immunogenicity analysis of the fusion protein[J];细胞与分子免疫学杂志;2005-05
【Co-citations】
Chinese Journal Full-text Database 8 Hits
1 LIU Bo;SUN Ze-qiang;LIU Qing-yong;CHEN Jie;WANG Si-jun;YANG Guang-xiao;WANG Quan-ying;School of Medicine of Shandong University;Department of Urinary Surgery,Ji'nan Central Hospital Affiliated to Shandong University;Xi'an Huaguang Biological Engineering Co.Ltd.;;Construction and prokaryotic expression of recombinant gene G250 antigenic peptide-HBcAg and the immunogenicity analysis of the fusion protein[J];西安交通大学学报(医学版);2014-01
2 FENG Gaifeng1,JIN Hui1,WANG Weixi1,QIAN Yihua1,WANG Quanying2,YANG Guangxiao2(1.Department of Human Anatomy and Histo/Embryology,School of Medicine,Xi’an Jiaotong University,Xi’an 710061;2.Xi’an Huaguang Biotechnology Company,Xi’an 710025,China).;Prokaryotic expression and immunogenicity of the chimeric HBcAg containing Aβ_(1-15)[J];中南大学学报(医学版);2012-03
3 FENG Gai-feng1,WANG Jun-yang2,JIN Hui1,WANG Wei-xi1,QIAN Yi-hua1,YANG Wei-na1,WANG Quan-ying3,YANG Guang-xiao3 1Department of Human Anatomy and Histo/Embryology,2Department of Microbiology and Immunology,School of Medicine,Xi'an Jiaotong University,Xi'an 710061;3Xi'an Huaguang Biotechnology Company,Xi'an 710025,China;Prokaryotic expression and immunogenicity analysis of the chimeric HBcAg containing APP beta cleavage site peptide and Aβ_(1-15)[J];细胞与分子免疫学杂志;2011-11
4 FENG Gai-feng,HU Hai-tao,JIN Hui,WANG Wei-xi,QIAN Yi-hua Department of Human Anatomy and Histoembryology,Xi'an Jiaotong University College of Medicine,Xi'an 710061,China;Immunization with recombinant HBcAg and β-amyloid peptide fusion protein promotes clearance of intrahippocampally injected β-amyloid peptide in rats[J];南方医科大学学报;2011-07
5 LIU Zhao-hui,FENG Gai-feng,LI Yue-ying,MA Yan-bing,HU Hai-tao△. Department of Anatomy,Histology,and Embryology at School of Medicine,Xi'an Jiaotong University,Key Laboratory of Environment and Genes Related to Disease,Ministry of Education,Xi'an 710061,China;Biological Effects of Immune Serum from Fusion Protein of β-amyloid Peptide and HbcAg/MIR on AD Transgeneic Cells[J];四川大学学报(医学版);2007-04
6 Hu Haitao,Dong Weijiang(Department of Anatomy and Histology & Embryology,Medical School of Xi'an Jiaotong University,Xi'an 710061,China);Recent progress and prospect of research on Alzheimer's disease[J];西安交通大学学报(医学版);2007-01
7 DUAN Xiao-yi~1, WANG Jian-sheng~1, GUO You-min~ 2* , HAN Jun-li~2, WANG Quan-ying~3, YANG Guang-xiao~3 ~1Department of Nuclear Medicine, the First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061; ~2Department of Image Center, the Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710004, China;Construction and prokaryotic expression of recombinant gene EGFRvIII-HBcAg and immunogenicity analysis of the fusion protein[J];细胞与分子免疫学杂志;2007-01
8 FENG Gai-Feng,HU Hai-Tao,JIN Hui,et al.Department of Human Anatomy and Histology/Embryology,Medical College of Xi'an Jiaotong University,Xi'an 710061,Shanxi,China;Sera of the mice immunized with fusion protein CAC inhibit fibrous formation of β-amyloid peptide[J];中国老年学杂志;2005-10
【Co-references】
Chinese Journal Full-text Database 2 Hits
1 DUAN Xiao-yi~1, WANG Jian-sheng~1, GUO You-min~ 2* , HAN Jun-li~2, WANG Quan-ying~3, YANG Guang-xiao~3 ~1Department of Nuclear Medicine, the First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061; ~2Department of Image Center, the Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710004, China;Construction and prokaryotic expression of recombinant gene EGFRvIII-HBcAg and immunogenicity analysis of the fusion protein[J];细胞与分子免疫学杂志;2007-01
2 FENG Gai-feng~1, HU Hai-tao~1, LI Yue-ying~1, HAN Hua~1, WANG Quan-ying~2, YANG Guang-xiao~2~1Department of Human Anatomy, Medical College, Xi'an Jiaotong University, Xi'an 710061; ~2Xi'an Huaguang Biotechnology Company, Xi'an 710025, China;Construction and prokaryotic expression of recombinant gene c-Aβ-c and the immunogenicity analysis of the fusion protein[J];细胞与分子免疫学杂志;2005-05
【Secondary Citations】
Chinese Journal Full-text Database 5 Hits
1 FENG Gai-feng~1, HU Hai-tao~1, DONG Wei-jiang~1, WANG Quan-ying~2, YANG Guang-xiao~2~1Department of Human Anatomy, Medical College, Xi’an Jiaotong University, Xi’an 710061; ~2Xi’an Huaguang Biotechnology Company, Xi’an 710025, China;Construction of recombinant plasmid pYTA/Aβ-HBcAg and its expression in E.coli[J];细胞与分子免疫学杂志;2004-06
2 Hu Haitao, Feng Gaifeng, Dong Weijiang, Wang Quanying, Yang Guangxiao (Department of Human Anatomy, Medical School of Xi'an Jiaotong University, Xi'an 710061, China);Prokaryotic expression of fusion gene Aβ-HBcAg and analysis of the immunoreactivity and immunogenicity of the expression protein[J];西安交通大学学报(医学版);2004-03
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