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ConstructionandVerificationofDoubleT-DNAPlantExpressionVectoroftheAt2Gene(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2012年12期
Page:
107-112
Research Field:
Publishing date:

Info

Title:
ConstructionandVerificationofDoubleT-DNAPlantExpressionVectoroftheAt2Gene
Author(s):
WANGWen-lingWANGXian-leiXiongLi-manGAOXing-wangLIGuan
CollegeofLifeScienceandTechnology,XinjiangUniversity,Urumqi,Xinjiang830046
Keywords:
At2genedoubleT-DNAplantexpressionvectormarker-free
PACS:
Q946
DOI:
-
Abstract:
Takingmeloncultivars‘MR-1’asmaterial,theenzymaticresistancegenesAt2wasamplifiedbyPCRmethodandtheBamHIenzymesiteandSacIenzymesitewasasoadded.Theresultsshowedthatitsaminoacidsequencehomologywas99%comparedwith“At2”genepublishedinGeneBankafterBLASTinNCBI;thenucleotidesequencehomologywas99.43%afteranalysisbyDNAMAN.TheacquiredgenewasligatedtoplasmidG-pPTN133,andthereforetheGUSgenewasreplacedofvectorA-pPTN133wasconstructed.TheplasmidA-pPTN133wasdigestedbySacIandthenligatedtothevectorpPTN133-whichwasasodigestedbySacI,sotheplantexpressionvectorAt2-pPTN133withdoubleT-DNAwasfinallyobtained.TheresultsshowedthatenzymesdigestionandPCRdemonstratedthecorrectnessofthevector.ThedoubleT-DNAvectorcanbetransformedintomelonmediatedbyAgrobacteriumtumufaciens.

References:

[1]HuangJ,RozelleS,PratC,etal.PlantBiotechnologyinChina[J].Science,2002,295:674-677.
[2]QianY.Analysisofadvantagesanddisadvantagesontransgeniccrops[J].BiotechnologyInformations,1999,5:7-11.
[3]董志峰,管华诗,马荣才,等.转基因植物中外源非目的基因片段的生物安全研究进展[J].植物学报,2011,43(7):661-672.
[4]雷茂良,程金根.全球转基因植物发展现状[J].生物技术通报,1988(6):30-32.
[5]Ow,DW,2011,Therightchemistryformarkergeneremoval[J].NatureBiotechnology,19(2):115-116.
[6]KomariT,HiheY,SaitoY,etal.VectorscarryingtwoseparateT-DNAsforco-transformationofhigherplantsmediatedbyAgrobacterium,tumefaciensandsegregationoftransformantsfreefromselectionmarkers[J].PlantJournal,1996,10(1):165-174.
[7]CleaveAP,MitradS,MudgeSR,etal.Selectablemarker-freetransgenicplantswithoutsexualcrossingtransientexpressionofcrerecombinaseanduseofaconditionallethaldominantgene[J].PlantMolecularBiology,1999,40(2):223-235.
[8]DaleEC,OwDW.Genetransferswithsubsequentremovaloftheselectiongenefromthehostgenome[J].ProcNatlAcadSciUSA,1991,88:10558-10562.
[9]GoldsbroughAP,LastrellaCN,YoderJI.ranspositionmediatedrepositioningandsubsequenteliminationofmarkergenesformtransgenictamato[J].NatureBiotechnology,1993,11:1286-1292.
[10]RusseellSH,HoopesJL,OdellJT.Directedexcisionofatransgeneformtheplantgenome[J].MolecularGeneticsandGenomics,1992,234:49-59.
[11]MikiB,MchughS.Selectablemarkergenesintransgenicplants:applications,alternativesandbiosafety[J].Biotechnology,2004,107(3):193-232.
[12]TalerD,GalperinM,BenjaminI,etal.PlanteRGenesthatencodephotorespiratoryenzymesconferresistanceagainstdisease[J].ThePlantCell,2004,16:172-184.
[13]ThomasCE,CohenY,McCreightYD,etal.InheritanceofresistancetodownymildewinCucumismelo[J].PlantDis,1988,72:33-35.
[14]XingA,ZhangZ,SatoS,etal.TheuseofthetwoT-DNAbinarysystemtoderivemarker-freetransgenicsoybeans[J].InVitroCell.Dev.Biol.-Plant,2000,36:456-463.
[15]萨姆布鲁克J,D.W.拉塞尔.分子克隆实验指南[M].黄培堂,译.3版,北京:科学出版社,2002.

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Last Update: 2014-09-19