Cloning Of VP3 Gene Of Goose Parvovirus And Construction Of Recombinant Fowlpox Virus Transfer Vector

The pathogen of goose plaque, goose parvovirus (GPV), can cause acute infectiousdisease characterized by acute intestitis and inflammation of liver, kidney and heart ingoslings and Muscovy duckings under 1 month of age, which threaten the goose industry.Goose plaque was first reported in China in l956, followed by severaI reportS of the diseaseand the virus isolation in many countries. The purPose of this study was to clone the majorstructural protein VP3 gene of GPV Hl isolate after PCR, and express by protokaryotic andeukaryotic exPressing system, then develop molecuIar diagnostic reagent of goose pIaque andconstruct recombillat fowlpox vina life vector vaccine.In this research, the GPV Hl isolate was propagated with 13 day's duck embryos. Apair of primers GFlGR used to arnplify VP3 gene was designed using Oligo4.I softwareaccording to the whole nucleotide sequence of GPV B isolate published by Zadori. The majorstructural protein VP3 gene was amplified from the DNA of GPV Hl isoIate by Polymerasechain reaction (PCR), and then cloned into pMDl8-T vecter. The recombinant plasmid wasidentit1ed with restriction endonuclease, PCR, then sequenced. The resuIt of sequenceanalysis showed that the VP3 gene is 1605bp and inc1uds a complete open reading frameencoding a protein of 534 amino acids. The Hl isolate shares 98.5% and 98.3% identity withB isolate at nucIeotide and amino acid levels respectively. 24 nucleotides differences existedbetween the VP3 gene of Hland B isolates are mainly distributed from 312bp to 4l7bp and79lbp to I276bp, the two regions might be mainly the high mutation regions. The deducedamino acid sequence of Hl isolate has nine amino acids differences with GPV B Jsolate whichlocates mainly in the region from 266aa to 417aa.VP3 gene of Hl isolate of goose parvovirus derived from recombinant pIasmid pPROEXHTb-VP3 was subcloned into EcoRl site of pSY538, and the reporter gene LacZ withpromoter pll was cloned into SmaI site of recombinant pIasmid. Both VP3 gene and LacZgene were cloned intO NotI site of pSY681, recombinan FPV transfer vector containing VP3gene of GPV was obtained. The result is basis of construction of recombinant FPV expressingVP3 gene of GPV and GPV genetic engineering vaccine.PostNe: TlAN LihOng Majort Preventive Veterineq ScienceAdvisort Pro f JIA YOngqing Pro f WANGjunwei...