Construction of Co-expressing Plasmid of FMDV P1-2A Gene with Porcine IFN-γ Gene and Expression in BHK-21 Cells

doi:10.7668/hbnxb.2010.05.002

Abstract

Abstract: To construct the co-expressing plasmid of FMDV P1-2A gene with porcine IFN-γ gene and express in BHK-21 cells. A P1-2A gene of foot-and-mouth disease virus(FMDV) was amplified by PCR from the plasmid pGEM-P12A and subcloned into pBudCE-IFN vector for sequencing after purification and digestion. A recombinant plasmid pBudCE-IFN-P1-2A was constructed and transformed into BHK-21 cells with liposome for in vitro expression. 48 hours after transfection,the mRNAs and expressed proteins of P1-2A and IFN-γ were detected by RT-PCR and direct immunofluorescence assay,respectively. The results showed that P1-2A and IFN-γ were successfully transcripted and expressed in BHK-21 cells. It was proved that the recombinant plasmid pBudCE-IFN-P1-2A was successfully constructed and expressed in vitro,providing a solid basis for development of DNA vaccine against FMDV

Key words: FMDV, P1-2A, IFN-γ, Co-expression

CLC Number:

WU Guohua, ZHANG Qiang, YAN Xinmin, HOU Junling, ZHAO Zhixun, CUI Lifan, LI Jian, ZHU Haixia, ZHU Caizhu. Construction of Co-expressing Plasmid of FMDV P1-2A Gene with Porcine IFN-γ Gene and Expression in BHK-21 Cells[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(5): 11-15. doi: 10.7668/hbnxb.2010.05.002.

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