Cloning and Expression of N and P of Human Respiratory Syncytial Virus RNA Polymerasa Complex

The n and p genes from subgroup A human respiratory syncytial virus (HRSV), were amplified by RT-PCR. The n product was cloned into pGEM-T easy vector and that of p into pcDNAII vector and the two genes were subcloned into the eukaryotic expression vector pcDNA3.1(+). The resulting recombinant plasmids pcDNA3.1(+)/N and pcDNA3.1(+)/P were checked by restriction enzymes and DNA sequencing and were tranfected into COS-7 cells with the aid of Lipofectamine 2000. Expression of N and P were detected by western blots at 72 hours post transfections. Expressed N and P in COS-7 cells was confirmed by western blot. The constructed expression vectors will be used for the further reverse genetics experimentations on HRSV.