Cloning and Construction of Eukaryotic Expression Vector of HN Gene of Newcastle Disease Virus Roakin Strain

To clone The HN gene of Newcastle disease virus(NDV) Roakin strain and construct a eukaryotic expression vector which can be expressed in Chinese Hamster Ovary Cells CHO/dhfr-.The HN gene of Newcastle disease virus(NDV) Roakin strain was amplified by RT-PCR and cloned into pMD18-T vector and sequenced.The HN gene of the virus and dhfr gene was cloned into high efficient eukaryotic vector pCI-neo.PCR amplification,restriction emzyme digestion and sequencing analysis was applied to identify the recombinant expression plasmids.The sequence homology of HN cloned with the HN(AY289000)on GenBank was 99.8%;Mammalian recombinant expression plasmid pCI-HN-D was constructed.Recombinant expression plasmid pCI-HN-D harboring dhfr gene and HN gene of NDV has been successfully constructed and lay the foundation of high-level expression of HN protein of NDV in mammalian cell CHO/dhfr-.