Tricistronic Plasmid Expressing the T7 RNA Polymerase and Measles Virus N and P Proteins for Rescue of Measles Virus AIK-C Strain

Up to now, several attenuated measles virus vaccine strains derived from the Edmonston B vaccine consisting of Schwarz/Moraten, Zagreb, and AIK-C have been introduced for the rescue of their relative viruses through reverse genetics. In most studies, the measles virus rescue was done by supplying a cell line that expresses T7 RNA polymerase and measles virus N and P proteins as accessory proteins. The present study aimed to evaluate the rescue eÿciency of the recombinant measles virus AIK-C vaccine strain by using a tricistronic expression plasmid. In this study, the rescue of the recombinant measles virus AIK-C vaccine strain was performed by co-transfection of three plasmids, including the cloned antigenomic cDNA of measles virus, a tricistronic expression plasmid that contained T7 RNA polymerase and measles virus N and P genes, and measles virus L polymerase expression plasmid. To increase the rescue eÿciency, the transfected HEK-293 cells were co-cultured with Vero cells. As a result, the use of tricistronic expression plasmid that concomitantly encoded three necessary genes for the rescue of the measles virus led to the viral cytopathic e ect with high eÿcacy five days post-transfection. Finally, the co-culture of transfected HEK-293 cells with Vero cells showed a relatively fast induction of viral cytopathic e ect.