CLONING AND SEQUENCING OF HAEMAGGLUTININ- NEURAMINIDASE (HN) GENE OF NEWCASTLE DISEASE VIRUS (NDV)
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Date
2008-11
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SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA
Abstract
ABSTRACT :
It is obvious to say that, the poultry farmer is the ultimate looser both in the farm of economy as well as loosing the chicken because of high mortality due to New- castle Disease (ND).
Since the virus is having pleurality in antigenecity with broad host range, it became impediment to develop a suitable vaccine for that geographical location. Hence a modest attempt had been made for the sequence information of HN gene of Indian vaccine strain K which would provide a valuable information for understanding molecular epidemiology, distribution, predicting NDV origin, genotyping, phylogenetic relationship, pathogenecity and virulence. Hence the molecular charecterization of K strain was very important with particular reference to HN gene.
The virus, that was cultivated in chicken embryos was subjected to RNA extraction by Trizol method and the nucleic acid was amplified by employing RT-PCR. The PCR amplicon was cloned into pDrive cloning vector and confirmed the gene size as 1731bp which coded for 577 amino acids .
The nucleotide and the deduced amino acid sequences of ten different isolates were multiple aligned with available database (GenBank) using Clustal W to find out the variations between different serotypes. The multiple alignment among these sequences indicated the
conserved nature of HN protein with respect to twelve cysteine residues and five glycosylysation sites . Subsequently, the phylogenetic relationship for these sequences was analysed using PHYLIP software. The phylogenetic tree analysis revealed correlation of K strain with other strains with varied percentage of homology.
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