ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2011, Vol. 42 ›› Issue (4): 521-526.doi:

• 预防兽医 • Previous Articles     Next Articles

Construction of Recombinant Vector of VP2 Gene of Foot-and-mouthDisease Virus and Establishment of Stable Cell Line for Its Stable Expression

XIE Jin-lu1,2,4, WANG Hong-mei2*, LIU Guo-yi1*, WU Jian-ming2,
LIU Xiao2, GAO Yun-dong2, YU Li3, ZHONG Ji-feng2, HE Hong-bin2*
  

  1. 1.Basic Medical College, Harbin Medical University, Harbin 150086, China;2. Dairy Cow Research Center, Shandong Academy of Agricultural Science,Jinan 250100, China; 3. Harbin Veterinary Research Institute, Harbin 150001, China; 4.Qinghuangdao Medical School, Qinghuangdao 066000, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-04-20 Published:2011-04-20

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.