关键词: 口蹄疫病毒, VP2, 转染, 稳定细胞系

Abstract: The aim of the study was to construct a recombinant vector, which including VP2 gene of Asia I strain of foot-and-mouth disease virus (FMDV), and to establish the BHK-21 cell line, which stably expressing VP2 gene. VP2 gene of FMDV was amplified from Asia I strain by RT-PCR, and its complete cDNA was cloned into pIRES2-EGFP vector. The recombinant pIRES2-EGFP-VP2 vector, which could express VP2 and EGFP proteins, was confirmed by sequencing analysis, the transient expression of VP2 in 293T cells transfected with the recombinant plasmid via LipofectamineTM 2000 was then determined by Western blot. Furthermore, BHK-21 cells were transfected with pIRES2-EGFP-VP2, and anti-G418 cell clones were screened using G418, both G418 and GFP positive cell clones were continually cultivated after GFP detection, their expressions of VP2 gene were confirmed by Western blot. The results showed that the recombinant plasmid pIRES2-EGFP-VP2 correctly encoded VP2 gene and transiently expressed in 293T cells- BHK-21 cells were transfected with recombinant plasmid, then BHK-21 cell clones expressing VP2 were obtained by G418 pressure selection, after continuous passage for 60 days, BHK-21 cell lines stably expressing VP2 gene of FMDV were established. The above results indicated that BHK-21 cell lines stably expressing VP2 gene of FMDV were established, and it provides a good platform to study the role of VP2 gene during FMDV induces apoptosis.

Key words: FMDV, VP2 gene, transfection, stable cell lines