7.5NS04CMV2a91.972.091.693.691.694.887.595.271.894.791.991.491.492.193.493.495.08
Thedown?leftisnucleotideidentities,theupper?rightisaminoacididentities.1-19arecorrespondingwiththesequenceNo.intheleftoffigurefromuptobottom
.
M:DL2000;1:CR2?BKsequence.Fig.3 PCRamplificationofCR2?B
K
M:DL4500;1:piRDCR2.SwithClaI.
Fig.5 Therestrictionenzymedigestionofrecombinantplas?
midpiRDCR2.
S
M:DL2000;1:CR2?BKsequence.
Fig.4 PCRamplificationofCR2?SB
ObtainingoftransgenictobaccoplantsTransgenictobaccoplantsDNAwasextracted,andthentheamplificationwascarriedoutbyprimersCR2Kp/CR2SB
,the1.2%ofagarosegelandCR2Bg/CR2SBrespectively
,itcouldbeconcludedelectrophoresiswasusedforanalysis
thatthePCR_CR2_BKfragmentwith693bpandthePCR_CR2_SBfragmentwith741bpwereamplified(Fig.7,8)
,
M:DL4500;1:pBi35SCR2withBamHⅠ.
Fig.6 Therestrictionenzymedigestionofrecombinantplas?
midpBi35SCR2
indicatingthattheobtainedtobaccoplantshadbeensuccess?
fullytransformedintothepBi35SCR2expressionvectorAgrobacterium?mediatedmethod.
72AgriculturalScience&TechnologyVol.11,No.5,2010
[10]
.Theconstructedexpres?theefficiencyofgenesilencing
icotianato?sionvectorpBi35SCR2wastransformedintotheN
baccowithAgrobacterium?mediatedmethod,theobtainedtransgenictobaccoplantscouldbeobservedontheantiviralabilityoftransgenictobaccoplantsbychallengetest,ifthe
,itcouldbefurtherappliedtotomatopro?effectwasobvious
cessing,whichhadlaidthefoundationfortheanti?CMVpro?cessingtomatoplants.
M:DL2000;1:pBi35SCR2vectorcontrol;2-5:pBi35SCR2vec?
tortransgenictobaccolines;6:PBI121transgenictobacco.Fig.7 PCRidentificationofCR2_BKfragmentintransgenicto?
baccoplant
s
References
[1]YANGS(,ZHANGGH(.Progressonresistancein?gh)ijk)
(heritanceofcucumbermosaicviruslmnopqprstuvw
[J].TianjinAgriculturalSciences(,2008,14z{|},-)xy)(2):55-58.[2]EAMENSA,WANGMB,SMITHNA,etal.RNAsilencingin
:yesterday,today,andtomorrow[J].PlantPhysiol,2008,plants147(2):456-468.[3]MATZKEM,MATZKEAJ,KOOTERJM.RNA:guidinggenesilen?
[J].Science,2001,293(5532):1080-1083.cing
[4]ZAMOREPD,TUSCHLT,SHARPPA,etal.RNAi:double?stran?
dedRNAdirectstheATP?dependentcleavageofmRNAat21to23
J].Cell,2000,101:25-33.nucleotideintervals[
[5]SIJENT,KOOTERJM.Post?transcriptionalgenesilencing:RNAs
ontheattackoronthedefense?[J].Bioessays,2000,22(6):520-531.[6]SHARPPA.RNAinterference[J].Genes&Development,2001,
15(5):485-490.[7]GUOXQ(,LUSE(,ZHUCX(,etal.~ ) ) )
PVY)inRNA?mediatedviralresistanceagainstpotatovirusY(
transgenictobaccoplants(NA Zrps r R
[J].ActaPhytopathologicaSinica( YpqZ ) Fp