Generation of a Vero Cell Line Stably Expressing African Swine Fever Virus P54 Protein

doi:10.13560/j.cnki.biotech.bull.1985.2019-1099

Abstract

Abstract: In order to establish the Vero cell lines that may stably express African swine fever virus（ASFV）,the fusion gene fragment from the ASFV-P54 gene with green fluorescent gene Azami Green was cloned into the lentivirus vector pLV-puro. The resulting plasmid,pLV-ASFV-P54-AG,was then co-transfected HEK-293V cells together with packaging plasmids pH1 and pH2 for packaging recombinant lentiviruses. The generated lentiviruses were then utilized to infect Vero cells mediated in the presence of polybrene. A Vero cell line,constitutively expressing the ASFV-P54 protein,was screened and designated as Vero-AG-ASFV-P54. Moreover,indirect immunofluorescence assays further demonstrated that the generated cell line reacted with P54 polyclonal antibody. The results further verified by National Veterinary Research Institute in Poland showed that the cell line reacted with the positive sera of ASFV antibody,but did not react with the negative sera. The above results reveal that the Vero-AG-ASFV-P54 cell line generated in this study may stably and efficiently express ASFV-P54 protein with biological activity.

Key words: African swine fever virus, P54, Azami Green, Vero cells, lentivirus vector

WANG Cai-xia, DU Fang-yuan, LIN Xiang-mei, Grzegorz Wozniakowski, WANG Qin, FENG Chun-yan, WU Shao-qiang. Generation of a Vero Cell Line Stably Expressing African Swine Fever Virus P54 Protein[J]. Biotechnology Bulletin, 2020, 36(5): 139-144.

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