CLONING OF HN GENE OF NEWCASTLE DISEASE VIRUS AND ITS EXPRESSION IN E. COLI

The HN gene with a length of 1 713 bp of Newcastle Disease Virus (NDV) was amplified by RT-PCR, the PCR product was purified and cloned into pGEM-T vector, and the recombinant plasmid was sequenced. According to the results of HN gene sequencing another pair of primer containing ECOR I restriction site was designed to amplify the HN fragment coding for the globular domain, the main functional region of HN protein. The HN fragment was then inserted at the 3' terminus of soc gene of pSOC plasmid, The recombinant plasmid pSOC-HN was used to transform E. coli BL-21(DE 3) to induce HN protein expression with 1 mmol/L IPTG. A SOC-HN fusion protein band with a molecular weight of 67 KD was detected on the SDS-PAGE gel and nitrocellulose membrane. The expression products can react specifically with antisera against Newcastle Disease Virus.