鵝細小病毒VP3基因克隆及重組禽痘病毒轉(zhuǎn)移載體的構(gòu)建.pdf

1、東北農(nóng)業(yè)大學(xué)碩士學(xué)位論文鵝細小病毒VP3基因克隆及重組禽痘病毒轉(zhuǎn)移載體的構(gòu)建姓名：田麗紅申請學(xué)位級別：碩士專業(yè)：預(yù)防獸醫(yī)學(xué)指導(dǎo)教師：賈永清王君偉2002.5.1堡主蘭垡堡蘭苧塑塵堡苧∑!!苧里塞墮墨蔓塑苧壟墮蘭壁堡塑堡塑塑壁堡苧查些查蘭CloningofVP3GeneofGooseParvovirusandConstructionofRecombinantFowlpoxVirusTransferVectorABSTRACTnlepath

2、ogenofgooseplaque，gooseparvovims(GPV)carlcauseacuteinfectiousdiseasecharacterizedbyacuteintestitisandinflammationofliverkidneyandheartingoslingsandMuscovyduckingsunderlmonthofage，whichthreatenthegooseindustryGooseplaqueW

3、aSfirstrepeltedinChinain1956，followedbyseveralreportsofthediseaseandthevirusisolationinmanycountriesThepurposeofthisstudywastoclonethemajorstructuralproteinVP3ganeofGPVH1isolateafterPCR，andexpressbyprotokaryoticandeukary

4、oticexpressingsystem，thendevelopmoleculardiagnosticreagentofgooseplaqueandconstlXlCtrecombinantfowlpoxVillBlifevectorvaccineInthisresearch。theGPVH1isolatew∞propagatedwith13day’SduckembryosApairofprimersGF／GRusedtoamplifr

5、VP3ganeWaSdesignedusingOli904lsoftwareaccordingtothewholenucleotidesequenceofGPVBisolatepublishedbyZadoriThemajorstructuralproteinVP3goneWaSamplifiedfromtheDNAofGPVHIisolatebyPolymerasechainreaction(PCR)andthenclonedinto

6、pMDl8TvecterTherecombinantptasmidWaSidentifiedwithrestrictionendonucleasePC＆tbensequenced111eresultofseouenceanalysisshowedthattheVP3goneis1605bpandincludsacompleteopenreadingflameencodingaproteinof534aminoacidsTheHlisol

7、ateshares985％and983％identitywithBisolateatnucleotideandaminoacidlevelsrespectively24nucleotidesdifferencesexistedbetweentheVP3geneofHlandBisolatesKremainlydistributedfrom312bpto417bpand791bptO】276bp，thetwegionsmightbemai

8、nlythehighmutationregionsThededucedaminoacidsequenceofHIisolatehasnineaminoacidsdifferenceswithGPVBisolatewhichlocatesmainlyintheregionfrom266aatO417anVP3geneofHIisolateofgooseparvovirusderivedfromrecombinantplasmidpPROE

9、XHTbVP3WassubelonedintoEcoRIsiteofpSY538andthe∞portergeneLacZwithpromoterpl】WasclonedjntoSmaIsiteofrecombinantplasmidBothVP3geneandLacZgenewereclonedintoNot[siteofpSY681，recombinantFPVtransfervectorcontainingVP3geneofGPV