| GPV (Goose parvovrius) has been classified by its morPhological, biochemical, andclut-Ure characteristics as a member of the parvovirus genus of the parvoviridae. lt is a highlyfatal disease of gos1ings and Muscovy ducklings, the typical pathological lesions was found indigest tract, specially in small intestine. The disease has broken out in several countries, andalI isolated GPV were similar in antigenicity. The disease was diagnosed mainly ac:cording tolts clinico-pathology and combining the serologica1 and virological methods, but most ofthose methods were time-consuming and laborious, which delay the cure, so it is necessary todevelop a rapid, simple and special diagnostic method. Prevention and cure of the Derzsy'sdisease depended on vaccine and antiserum, antibodies of eggs.The vaccines includes gooseembryo and duck embryo vaccines which were used 1n breed goose and goslings, and thosevaccines have great effect in breeding goose, but the entire virion live vaccines and attenuatedvaccines exist many deficiencies. such as preclinical infection, dissemination of virus,recovery of viru5, etc. Those proplems can be sol\/ed by producing genetic engineering'asclne.GPV contins three capsid protein. VP1. VP2 and VP3.The genes, encoding them, werelocated in a same ORF, and they possess of common stop codons. Furthermore, amino acidsequence of VPl contained the fu1l sequence of VP2 and VP3, so VP1 gene become interestgene. In this research. a pair of primers were designed to amplify VPl gene by PCR accordingto the published sequence of GPV B strain in Genbank. The product of PCR was 2.2 Kb. Afteridentification of PCR product by restriction endonuclease the interest gene was inserted in theTha I. Tho I multiple cloning sites of prokaryotlc expression vector pPRoEXHTb toconstruct the recombinant prokaryotic expression vector GPl -pPRO of VP 1 of GPV strain H 1.The result of sequencing showed that VP1 included 2l99 bases, which encoded,732 amnioacids. The interest gene shared 98.5% and 93. l 8% in bases comparisoning with GPV strains Band YG, and 98.09%, 95.77% homoIogy in amino acid.The interest gene was inserted in the -Tha l. Tho 1 multiple cloning sites of donorplasimd pFAsTBAcHTb of baculovirus expression system. After analysis by restrictionendonuclease and PCR, the recombinant donor plasmid GPl-FAST was transforn1ed to thecompetent celI DHI0BAc which contalns the bacmid and the helper plasmid, the recombinantbacmid GPl -BAC was acquired which would express the VPl of GPV strain H l.The secondary structUre and antigenicity of VP 1, VP2, VP3 of GPV strain H l and VP1s25fot%&&Z Abstract $jhAAk#of GPV strains B, YG were predicted and comparisoned by Antheprot5.0 software. The resultsuggested that there was no difference between VPl, VP2 and VP3 of GPV HIstrain insecondny structure and antigenicity, but VPls of GPV strains Hl, B, YG were different inthose two respects. Comparison of VPl of GPV strain Hl with other parvoviruses showed thatthe homology between GPV strain H1 and AAV-2 was higher than that of GPV stains H1 andother aUtonomous parvoviruses.This study provided the basis for the further research about GPV molecular biologicalproperties and the physical, chemical character and biological activity of VPl. lt was preparedwork for the development of diagnostic reagent and genetic engineering vaccine. And it alsoprovided the evidence for taxonomy of GPV and its relationship in evolution.Postgraduaef Bo Ma Major f Preventive veterinary scienceAdvisor f Pro f Jun wei Wang... |
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