Construction of an infectious clone of Zika virus stably expressing an EGFP marker in a eukaryotic expression system

Virology Journal

Jintai Cai¹  Jing Gao¹  Weizhi Lu¹  Linjuan Shi¹  Jiayi Chen¹  Wei Zhao²  Bao Zhang² 

[1] Biosafety Level 3 Laboratory, School of Public Health, Southern Medical University, 510515, Guangzhou, China;Biosafety Level 3 Laboratory, School of Public Health, Southern Medical University, 510515, Guangzhou, China;Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, 510515, Guangzhou, China;

关键词: Zika virus;    EGFP marker;    Recombinant virus;    Eukaryotic expression;   

DOI  :  10.1186/s12985-021-01622-z

来源: Springer

【 摘 要 】

BackgroundZika virus is becoming one of the most widely transmitted arboviruses in the world. Development of antiviral inhibitor and vaccine requires an experimental system that allows rapid monitoring of the virus infection. This is achievable with a reverse genetic system. In this study, we constructed an infectious clone for Zika virus that stably expressing EGFP.MethodsA PCR-mediated recombination approach was used to assemble the full-length Zika virus genome containing the CMV promoter, intron, EGFP, hepatitis delta virus ribozyme, and SV40 terminator sequence for cloning into the pBAC11 vector to produce recombinant pBAC-ZIKA-EGFP. ZIKA-EGFP virus was rescued by transfection of pBAC-ZIKA-EGFP into 293T cells. The characterization of ZIKA-EGFP virus was determined by qPCR, plaque assay, CCK-8, and Western blot.ResultsRescued ZIKA-EGFP virus exhibited stable replication for at least five generations in tissue culture. ZIKA-EGFP can effectively infect C6/36, SH-SY5Y and Vero cells, and cause cytopathic effects on SH-SY5Y and Vero cells. The inhibition of ZIKA-EGFP by NF-κB inhibitor, caffeic acid phenethyl ester was observed by fluorescence microscopy.ConclusionOur results suggested that Zika virus infectious clone with an EGFP marker retained it infectivity as wide-type Zika virus which could be used for drugs screening.

【 授权许可】

CC BY