口蹄疫病毒VP1基因重组慢病毒载体的构建及VP1基因稳定表达细胞系的建立

doi:10.3864/j.issn.0578-1752.2010.16.023

Scientia Agricultura Sinica ›› 2010, Vol. 43 ›› Issue (16): 3455-3460 .doi: 10.3864/j.issn.0578-1752.2010.16.023

• VETERINARY SCIENCE • Previous Articles Next Articles

Construction of VP1 Recombinant Lentivirus Vector and Establishment of BHK-21 Cell Lines Stably Expressing VP1 Gene of FMDV

DAI Wen-jun, WANG Hong-mei, LIU Xiao, GAO Yun-dong, YU Li, WANG Li-qun, ZHONG Ji-feng, HE Hong-bin

(山东省农业科学院奶牛研究中心)

Received:2010-01-22 Revised:2010-04-05 Online:2010-08-15 Published:2010-08-15
Contact: HE Hong-bin，WANG Hong-mei,ZHONG Ji-feng

Abstract

Abstract:

【Objective】 To construct the lentivirus vector containing VP1 gene and to establish the cell line with stable expression of VP1 gene of foot-and-mouth disease virus (FMDV). 【Method】 The full-length VP1 gene was amplified by RT-PCR, and VP1 gene was cloned into FG9 vector, then recombinant vector was confirmed by restricting enzyme digestion and DNA sequence. The recombinant plasmid was transfected into BHK-21 cells through LipofectamineTM 2000, and the GFP positive cells were screened via FACS after 96h. The expressions of VP1 gene were confirmed by Western-blot. 【Result】 FG9-VP1 was constructed successfully and the VP1 gene could be stably expressed in BHK-21 cel1s. 【Conclusion】 The recombinant lentivirus vector containing VP1 gene was cloned successfully and the stable cell line expressing the VP1 gene was established.

Key words: FMDV, VP1 gene, transfection, stable cell lines

DAI Wen-jun,WANG Hong-mei,LIU Xiao,GAO Yun-dong,YU Li,WANG Li-qun,ZHONG Ji-feng,HE Hong-bin
. Construction of VP1 Recombinant Lentivirus Vector and Establishment of BHK-21 Cell Lines Stably Expressing VP1 Gene of FMDV
[J].Scientia Agricultura Sinica, 2010, 43(16): 3455-3460 .

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