抗黄瓜花叶病毒RNAi载体的构建及烟草的转化(4)

发布时间:2021-06-05

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

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

,2001,31(4):349-356. -d)

[8]NIUYB(,QINGL(,ZHOUXP(.Mechanism ) )3 )

(RNA ¡ofRNAsilencinganditsapplicationforvirusresistance

)[J].ChinaBiotechnology(¢£¤¥rpq¦ <=YF§¨©

,2004,24(2):76-79.ª)

[9]LIF(,ZHOUXP(,LIDB(,etal.cDNAclo«¬)3 )«­®)

ningandsequenceanalysisofcoatproteingenesofTMVandCMVisolatedfromtobaccoinYunnanprovince(¯P° nopq]l

[J].JournalofZhejiangmnopq±²³´ Nµ¤¶·[¸)

:Agric&LifeSci(AgriculturalUniversity¹ºc--d:|}»Y¼,

,2000,26(3):261-265.-1)

[10]BRUMMELKAMPTR,BERNARDSR,AGAMIR.Asystemfor

stableexpressionofshortinterferingRNAsinmammaliancells[J].Science,2002,296(5567):550-553.

M:DL2000;1:pBi35SCR2vectorcontrol;2-5:pBi35SCR2vec

tortransgenictobaccolines;6:PBI121transgenictobacco.Fig.8 PCRidentificationofCR2_SBfragmentintransgenicto

baccoplants

Discussions

TheCMVRAN2genomesequenceofCMVNS04isolates

,andthefunctionalgenesequenceawasclonedinthisstudy

lignmentandphylogeneticanalysisshowedthatthecucumbermosaicvirusCMVNS04isolatesofXinjianghadahighernucleotideandaminoacidhomologywithDQ412731isolates

,Zhejiang,whichreached98.0%and96.5%refromChina

spectively.Inthisstudy,theconservedsequencesinCMVRNA2genomewhichwaslessstudiedwasselectedasthetar

,andvectorwouldformhpRNAgetsequenceofRNAivector

structureaftertranscription,andthehpRNAstructureincluded

,andtherewasaringstructureofasenseandantisensearms

about50bpbetweenthetwoarms,whichhelpedtoimprove

Responsibleeditor:LITingting

Responsibleproofreader:WUXiaoyan

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NAi摘要)[&'\]^_R`a1bcUde1fg(

张瑜,郑银英,赵峰玉,乔亚红,崔百明,向本春 (ãä+X%åæ3 z¦§¨©,ãä+X%3 %&,âçãä

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[NAi41]èéê>?ë{ìíRîï,«Õîïðñòó。[T-PCRõÂ,S04NA2MVRAN256]ô Rö÷>?ë{ìíNø$ùú¢ª 5Rûüý5ÐÑ。ÈþCûüýs5ÿ!"#$ L

Bi35SCR2CR5õÂÃÄîï ,%ÐÑ,èép¶&º'îï,«=º'îï()¬G;½¾åZ4*+5õÂÕº'îïðñòó, P

ðñ。[RNA2MV2aQ412731¢ª ~6w&78n9û87 :,78]-. #¸¢Ò/ º0,s12C5ÐÑ s3455D¢;'¼98.0%[96.5%;PCR/ º0,Bi35SCR2<¨=>èéÜp¶&º'îï,«=>Õº'îïðñòó。[],79<¨É»5ðûüòóÊ L?@Aí<¨5B «L ø$ùúê>?ë{ìíCGÜûD。

RNAi:;< >?ë{ìí;7 :;îï;òó;ð#

2008DFA30560);973计划前期研究专项(2008CB117018);石河子大学高层次人才科研启动项目(RCZX200732)。XYZ4 国际科技合作项目(

1984-),女,黑龙江哈尔滨人,硕士研究生,研究方向:植物基因工程。通讯作者,教授,博士生导师。=>?@ 张瑜(0100526  DEBC 20100629-ABC 2

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