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Expression of IV-AIV/AIV-IV Hemagglutinin Fusion Protein in E.coli

Author: HanZuoQi
Tutor: ShengJun
School: Jilin University
Course: Microbial and Biochemical Pharmacy
Keywords: recombinant Hemagglutinin protein 7ZTS/H1A1H5A2 7ZTS/H5A1H1A2
CLC: R373
Type: Master's thesis
Year: 2007
Downloads: 93
Quote: 0
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Abstract


Objective : To construct recombinant vector 7ZTS/H1A1H5A2 and 7ZTS/H5A1H1A2 and express them in Ecoli , detect the biological characteristics of recombinant protein product Hemagglutinin of IV/AIV and AIV/IV .Method:Fresh allantoic fluid from 10 day-old chicken embryos infected by H1N1 bird flu virus was used to obtain gene H1 by RT-PCR . A series of PCR reactions was used to get gene H1A1 and H1A2, and gene H5A1 and H5A2 from gene H1 and vector pMD18-T/H5. SOE PCR was used to connect each of them to become gene H1A1H5A2 and H5A1H1A2 which were connected with vector 7ZTS respectively. Then recombinant vector 7ZTS/H1A1H5A2 and 7ZTS/H5A1H1A2 was obtained and transfected into E.coli JM109 cell which was induced by IPTG at 37℃. Electrophoresis and western blot was used to identify the protein.Result:Sequencing and digesting analyse result was right. Electrophoresis showed the two kinds of recombinant protein were expressed well. Western blot analyse showed recombinant Hemagglutinin IV/AIV connected anti-H1 specifically recombinant Hemagglutinin AIV/IV connected anti-H5 specifically.Conclusion:Gene H1A1H5A2 and gene H5A1H1A2 were successfully obtained. Recombinant vector 7ZTS/H1A1H5A2 and 7ZTS/H5A1H1A2 were successfully constructed. Recombinant Hemagglutinin protein was expressed well and conserved its antigenic on its heavy chain.

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CLC: > Medicine, health > Basic Medical > Medical Microbiology ( pathogenic bacteriology,pathogenic microbiology ) > Human Virology ( pathogenic virus)
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