Expression of Prokaryotic Gene in Mammalian Cells Based on Recombinant Vaccinia Virus Encoding T7 RNA Polymerase

The vaccinia virus/T7 RNA polymerase transient expression system relying on the bacteriophage T7 RNA polymerase with several key advantages was widely used for protein expression.VP6-encoding cDNA of rotavirus strain TB-Chen was cloned and constructed by inserting into prokaryotic expression plasmid pETL between bacteriophage T7 promoter and terminator regions.When eukaryotic cells were transfected with the recombinant plasmid containing the VP6 gene and infected with recombinant vaccinia virus vTF7-3 which provides bacteriophage T7 RNA polymerase,the VP6 gene was expressed.The findings showed that cytopathic effects by vaccinia virus infection were fast and severe leading to low level of protein expression.If cytopathic effects of the vaccinia virus can be reduced,higher level of the protein might be obtained.The findings provided a helpful means to further study the VP6 function and utilize the vaccinia virus/T7 RNA polymerase transient expression system.