Expression of 3C Proteinase of Type O Foot and Mouth Disease Virus in Mammalian Cells

doi:10.11924/j.issn.1000-6850.2013-3399

Abstract

Abstract:

To construct the eukaryotic expression plasmid of tpye O foot-and-mouth disaese virus(FMDV) 3C proteinase, and to express in BHK-21 cells, 3C gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from type O FMDV strain and then cloned into the eukaryotic expression vector pcDNA3.1(-) using the restriction enzyme XhoⅠand Hind Ⅲ, the recombinant plasmid pcDNA3.1(-)-3C was transfected into the BHK-21 cells by Lipofectamine ^TM 2000 Reagent. After the confirmation by PCR, enzyme digestion and sequence analyses, the resulting plasmid pcDNA3.1(-)-3C was constructed correctly; Western-blot analyses showed that the plasmid pcDNA3.1(-)-3C was successfully expressed type O FMDV 3C proteinase in BHK-21 cells. The construction of the eukaryotic expression plasmid would help to study the assembly of FMDV empty capsid in vitro and establish a basic experiment to develop the type O FMDV empty capsid vaccine in future.

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