Establishment and characterization of the reporter cell lines for varicella-zoster virus / 中华微生物学和免疫学杂志

ABSTRACT

Objective To establish the reporter cell lines for varicella-zoster virus(VZV)with ORF9G,the shortest and efficient sequence of the promoter for VZV ORF9,and ORF61F,the shortest and efficient sequence of the promoter for ORF61,and to characterize the cell lines.Methods The tandem promoters.T9G and T6lF,which were resulted respectively from the linkage of ORF9 in duplicate and of ORF6lF in duplicate.were cloned respectively into an individual reporter plasmid pGL3-basic.In this way,two recombinant promoter-reporter plasmids.pGL-T9G and pGL-T61F were constructed,in which the expression of reporter gene firefly luciferase was under the control of the upstream T9G or T61F.Along with the G418-resistant plasmid pCMV-script.the pGL-T9G and pGL-T6lF were respectively transformed into an in dividual Me Wo cell line.The grown G418-resistant cell clones were collected,and their firefly luciferase expressions post VZV infection was assayed.The best cell clones that have high firefly luciferase activity were chosen as reporter cell lines for VZV,of which the sensitivity and specificity were characterized. Results The activity of T9G or T61 F was two-fold as that of 9G or 61F.Two reporter cell lines,MV9G containing ORF9 ptomoter and MV6lF containing ORF61 promoter,were established successfully.Both cell lines showed fast.sensitive and specific response to VZV infection in a dose-dependent manner although the sensitivitv of MV9G Was somewhat higher than that of MV61F.Conclusion Each of both reporter cell lines for VZV may serve as a sensitive and specific research tool for further study especially on virus entry and antivi ral mechanisms.