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Zhang Y.,Nattonat Institute of Viral Disease Control and Prevention | Yu Z.,Nattonat Institute of Viral Disease Control and Prevention | Xin L.,Nattonat Institute of Viral Disease Control and Prevention | Chen Y.,Nattonat Institute of Viral Disease Control and Prevention | And 4 more authors.
Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology | Year: 2010

On the basis of successful cloning the full length hemagglulinin (HA) and neuramidinase (NA) gene and sequence analysis of influenza virus H1N1, part of the gene was ligated into pMETA. Expression vectors pMETA/HA (52-1 557 bp) and pMETA/NA (121-1 263 bp) were constructed and expressed in pMAD16 induced by methanol. Recombinant protein was purified through Ni 2+ affinity chromatography. Western blotting and ELISA were used to determine the antigenic activity of the recombinant protein. SDS-PAGE showed that the recombinant capsid gene could be overexpressed in Pichia methanolica. ELISA and Western blotting showed that the recombinant protein had antigenicity. ©2010 CJB, All right reserved.


PubMed | Nattonat Institute of Viral Disease Control and Prevention
Type: Journal Article | Journal: Sheng wu gong cheng xue bao = Chinese journal of biotechnology | Year: 2010

On the basis of successful cloning the full length hemagglulinin (HA) and neuramidinase (NA) gene and sequence analysis of influenza virus H1N1, part of the gene was ligated into pMETA. Expression vectors pMETA/HA (52-1 557 bp) and pMETA/NA (121-1 263 bp) were constructed and expressed in pMAD16 induced by methanol. Recombinant protein was purified through Ni2+ affinity chromatography. Western blotting and ELISA were used to determine the antigenic activity of the recombinant protein. SDS-PAGE showed that the recombinant capsid gene could be overexpressed in Pichia methanolica. ELISA and Western blotting showed that the recombinant protein had antigenicity.

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