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Citation: YU Jie-mei, He Jin-sheng*, LI Dong-liang, YANG Bing, YUAN Yuan. Cloning and Expression of N and P of Human Respiratory Syncytial Virus RNA Polymerasa Complex [J].VIROLOGICA SINICA, 2006, 21(3) : 213-216.

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.

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

    • 1. Department of Immunology, Anh.ui Medical University, Hefei 230032, China

    Abstract: 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.

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