Construction of pEGFP-3D Eukaryotic Expression Vector of FMDV 3D Gene and Expression and Localization of Fusion Protein Molecule in BHK-21 Cells

doi:10.7668/hbnxb.2008.04.002

Abstract

Abstract: To construct pEGFP -3D expression vector and to detect its transient expression and its localization in transfected BHK -211Depending on tow steps of cleavage with BamH I and Hind FMDV 3Dgene can be cleaved from pGEM -T -3D and inserted into the plasmid pEGFP -N11The eukaryotic expression vector pEGFP -3D was constructed, which can express 3D fusion protein. The recombinant plasmid pEGFP -3D was transfected into BHK -21 cells1The pEGFP -3D plasmid was identified by restircted enzyme analysis and DNA sequencing and it was showed it contained. the full length of 3D gene. Immunohistochemistry analysis showed that 3D -GFP fusion protein mainly localized in BHCK -21 cell nucleus1The 3D -GFP fusion protein expressed could react with FMDV polyclonal antibodies inwestern blot analysis, demonstrating that 3D -GFP fusion protein having an immunological activity structure. The pEGFP -3D expression vector makes it possible to study the expression of 3D -GFP fusion protein and the protein localization in transfected cells1Moreover, as a useful tool to study the bio logical function of 3D protein will to necessary.

Key words: Foo- t and -mouth disease virus, pEGFP -3D transient expression vector, Immunohistochemistry analysis, Immunological activity

CLC Number:

S432.4+

BI Yan-li, SHEN Xiao-yan, CONG Guo-zheng, LIU Xiang-tao, CHANG Hui-yun, CAI Xue-peng. Construction of pEGFP-3D Eukaryotic Expression Vector of FMDV 3D Gene and Expression and Localization of Fusion Protein Molecule in BHK-21 Cells[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(4): 5-9. doi: 10.7668/hbnxb.2008.04.002.

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