Screening of Rice Genes Interacting with p5b of Rice Black- Streaked Dwarf Virus

(整期优先)网络出版时间:2013-03-13
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Riceblack-streakeddwarfvirus(RBSDV)isarecognizedmemberofthegenusFijivirus,familyReoviridae.Itsgenomehastendouble-strandedRNA(dsRNA)segments(S1-S10),inwhichthefifthgenomesegment(S5)containstwoopenreadingframes(ORFs)withapartiallyoverlappingregion.ThesecondORFofRBSDVS5encodesaviralnonstructuralproteinnamedp5bwithunknownfunction.Torevealthefunctionofp5b,itsgenewasligatedintothebaitplasmidpGBKT7andanexpressionlibrarycontainingricecDNAswasconstructedusingplasmidpGADT7foryeasttwo-hybridassay.Thebaitproteinp5bwasdetectedinyeastbywesternblot,andtheresultofanauto-activationtestshowedthatp5bcouldnotautonomouslyactivatetheexpressionofreportergenesinyeast.Thenthebaitproteinp5bwasusedforscreeningthecDNAexpressionlibrariesofrice.Genefragmentsofsomepivotalenzymesinvolvedinphotosynthesis,respirationandotherimportantmetabolicprocesses,wereidentifiedtointeractwithp5binyeast,suggestingthattheseinteractionsmayplayrolesinsymptomdevelopmentininfectedplants.