抗黄瓜花叶病毒RNAi载体的构建及烟草的转化
时间:2025-03-12
时间:2025-03-12
AgriculturalScience&Technology,2010,11(5):69-72Copyright2010,InformationInstituteofHAAS.Allrightsreserved.
PlantProtection
ConstructionofRNAiVectorWhichResistCucumberMosaicVirusandTransformationofTobacco
ZHANGYu,ZHENGYinying,ZHAOFengyu,QIAOYahong,CUIBaiming,XIANGBenchun
KeyLaboratoryofAgricultrureBiotechnologyofShiheziUniversity/CollegeofLifeSciences,ShiheziUniversity,Shihezi832003
Abstract [Objective]AimedtoconstructRNAivectorresistanttocucumbermosaicvirusandtransferredthisvectorintotobacco.[Method]RTPCRmethodwasusedtoamplifycucumbermosaicvirusNS04andprocessRNA2genesequenoftomatoisolates.TheanalysisresultsofphylogenetictreedemonstratedthatthesequenceinRNA2encodedCMV2ahad98.0%and96.5%homologywithnucleotideandaminoacidof
,China.ThereplicasefragmentinCMVRAN2genewastakenastargetsequencetoconstructpBi35SCR2eukaryDQ412731isolateofZhejiang
oticexpressionvector,thentheexpressionvectorwasidentified.Throughagrobacteriummediatedmethod,theexpressionvectorwastransferredintotabaccoandPCRmethodwasusedtocheckthetransfer.ThePCRresultsdemonstratedthattheexperimenthadsuccessfullyconstructeukaryoticexpressionvectorofpBi35SCR2andtheexpressionvectorwassuccessfullytransferredintotabacco.[Conclusion]Theobtainedtransgenictobaccocouldbeusedaschallengetestmaterialinfollowingexperimentandprovidedfoundationforstudyingprocessingtomatoresistcucumbermosaicvirus.Keywords Cucumbermosaicvirus;Homology;RNAivector;Tobacco;Transformation
Cucumbermosaicvirus(CMV)isanimportantpathogentocauseyieldreductionandevennoharvestofavarietyofeconomiccrops,butthetraditionalantivirusbreeding,tissuedetoxificationandpesticidepreventionmethodscauseenvi
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ronmentalpollution,andtimeconsuming.Inrecentyears,thediscoveredphenomenonofgenesilencingisacommonphenomenonexistedintheprocessofbiologicallife,whichisanimportantwayoftheregulationofgeneexpression.ThegenesilencingofGMspecificcanbetransmittedbygraftingfromsilentstocktononsilentscion,thenthephenomenonofthetransferringofthiskindofgenesile …… 此处隐藏:2791字,全部文档内容请下载后查看。喜欢就下载吧 ……