Recombinant adenovirus-mediated transfer of the human rotavirus capsid protein gene Wa VP7sc into the human embryonic kydney [sic] cell line 293

Adenovirus-mediated expression of a modified simian rotavirus capsid protein (SA-11 VP7sc) in 293 cells has been demonstrated. Based on this observation and due to the high-level similarity between VP7 proteins from different rotavirus serotypes, VP7sc from the Wa strain (serotype Gl) was rescued into the adenovirus (Ad5) El region to obtain its expression following infection of 293 cell This gene which has been constructed and expressed in COS cells, was prepared from pJCWaVPTsc plasmid. The gene was first subcloned into Bluescript to facilitate selection of recombinants following ligation of the CMV promoter DNA upstream of it. This cassette was then subcloned into the polycloning site of pXCX3, a plasmid containing 16% left-hand sequences of Ad5. This step produced pXCVPV Plasmid which could give rise to the rescue of WaVP7 in the El region of Ad5 following cotransfection and homologous recombination with pJMlT, a plasmid containing aknost the entire genome of Ad5.