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| Construction and Biologic Activity Detection of Anti-Human Red Blood Cell ScFv-PRV gE Bifunctional Fusion Protein |
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Abstract Construction of bifunctional fusion protein which can conjugate both human red blood cell and antibody against PRV.2E8 gene(ScFv gene against H antigen of human erythrocytes)and kgE gene (the major epitope domain of glycoprotein gE of pseudorabies virus) ,amplified from the recombinant plasmid PMD-2E8ScFv and PMD-gE,respectvely,were spliced by overlaped extension PCR and the assembled bifunctional molecule was named 2E8kgE.The 2E8kgE digested with BamHI and EcoRI was inserted into pET-Trx vector,to yield the recombinan vector pET-Trx-2E8kgE.Then the recombinan vector pET-Trx-2E8kgE was transformed into BL21plysS and protein expressions was induced with IPTG.SDS-PAGE and western-blot analyses showed that the fusion peotein was 60.1kDa and the protein was specific to antisera against PRV. Erythrocyte agglutination test results indicated that 2E8kgE fusion protein can conjugate both human red blood cell and antibody against PRV.
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Received: 04 May 2009
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