Expression of Bt Gene in a Dicot Plant Under Promo.pdf
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Expression of Bt Gene in a Dicot Plant Under Promo.pdf

ExpressionofBtGeneinaDicotPlantUnderPromoter.pdf

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PakistanJournalofBiologicalSciences4(12):1518-1522,2001©AsianNetworkforScientificInformation2001ExpressionofBtGeneinaDicotPlantUnderPromoterDerivedfromaMonocotPlantM.A.Khan,R.Makhdoom,T.Husnain,M.Z.Saleem,K.Malik,Z.Latif,1I.AltosaarandS.RiazuddinCenterforAppliedMolecularBiology,UniversityofthePunjab,87-WestCanalBankRoad,ThokerNiazBaig-57300,Lahore,Pakistan1DepartmentofBiochemistry,UniversityofOttawa,40MarieCuriePrivate,Ottawa,Ontario,K1N6N5,CanadaAbstract:Thepossibilityofusingamonocot(maize)derivedubiquitin(Ubi)promotertoexpressafullymodifiedinsecticidalcrygeneofBacillusthuringiensisindicotplant,tobaccowasstudied.Tobacco(Nicotianatabacumcv.xanthi)plantsweretransformedwithBtgeneCry-1AcdrivenbyUbipromoter.Transgenicplantswereconfirmedfortransformation,geneexpressionandinsecticidalactivitythroughPCR,GUSassays,Southernblot,Westernblotanalysesandinsectbioassays.Bioassayswithcry-1ActransformedToandT1transgenicplantsshowedhighleveloftoxicitytowardsAmericanbollworm(Heliothisarmigera)giving100%mortalityofthelarvae.ThispaperreportedthatmonocotderivedUbipromoterexpressesaBtgeneinadicotplantinaneffectivemannertorenderthetransformedplantshighlyresistantagainstHeliothisarmigera.Keywords:Ubiquitinpromoter,tobacco,cry-1Acgene,Heliothisarmigera,Americanbollworm.IntroductionConstructionofanAgrobacteriumbasedplantexpressionTheimprovementintransformationtechnologyhasresultedinvectorcontainingcry-1AcgeneunderUbipromoter:ThethesuccessfulproductionofanumberoftransgenicplantCry1Acgenefragment,alongwithubiquitinpromoter(Ubipro)species.Insectresistanttransgeniccropscanbeproducedandnopalinesynthaseterminator(noster)wasexcisedfromwhichcanresultinimprovedyields,environmentalsafetyandplasmidUbi-Cry1Ac(Sardanaetal.,1996)withHindIIIpesticide-freeproduce.Therapidlyemergingfieldofplantenzyme.This~4kbfragmentwasligatedintoanbiotechnologyhasshownsubstantialprogressintheAgrobacteriumbasedplanttransformationvectorpKHG4(LedevelopmentoftransgenicplantsexpressingBtgenes,andG