高賴氨酸蛋白基因在轉(zhuǎn)基因生菜中的表達和遺傳研究.pdf

1、甘肅農(nóng)業(yè)大學碩士學位論文高賴氨酸蛋白基因在轉(zhuǎn)基因生菜中的表達和遺傳研究姓名：李興濤申請學位級別：碩士專業(yè)：作物遺傳育種指導教師：張金文劉玲20050601AbstraetVariousfactorsinfluencingtissuecultureoflettuce(LactucasativaL)wereinvestigatedandatransformationsystemwasestablishedwithexplantTransge

2、niclettucewasobtainedbyAgrobacteriumtumefaciecsmediatedtransformationofleafdiscLysinerichproteingenewasintroducedintolettuce，TheresultsindicatedthattheintroducedgenewasintegratedintotheTOandTlgenerationtransgenicplantsAn

3、efficientgenetransfersystemforlettucewasestablishedExplantswerefirstpre—culturedonMSmediumfor1—2days，thenco—culturedonoriginalMSmediumwithanAgrobacteriumtumefaciensstrain(LBA4404)carryingthemodifiedpBll21plasmidwhichcont

4、ainslysine～richproteingenedrivenbyCaMV35SpromoterExplantweretransferredtoMSmediumwithKan100m【g／LandCarb500mg／L，andbudselongatedonMSmediumcontainingthesameconcentrationKanAllelongatedshootrootedon112MSmediumthentransferre

5、dshootgrew2cmStudiesontheeffectsofvariousfactorsinfluencingtransformationindicatedthataseriousoffactors，includingtheageofexplant，theconditionofAgrobacteriumtumefaciens，thepre—cultureofexplants，thetimeoftransformationande

6、o—culture，influencestherateoftransformationoflettuceTheresultsindicatedthat：theoptimalexplantsagefortransformationwas23days；theAgrobacteriumtumefaciensinthelogarithmtimehavethemostabilityfortransformation；theoptimaltimeo

7、ftransformationwas20minwhentheAgrobacteriumtumefacienswasdilutedtoOD600208：theoptimaldurationofpre—culturewas1～2daysthatwouldimprovetherateoftransformation；Thedurationof23daysco—culturemightimprovetherateoftransformation

8、tooThetransgenicnatureofregenetantswasindicatedbyplantgrowthonmediumcontainingKanPCRandsouthernblotanalysisdemonstratedfurtherlysine—richproteingeneintegratedintotransgenicplantsSimultaneouslytheresultofRTPCRdetectioncon

9、firmedthetargetgenehasbeenexpressedontheleveloftranscriptionDataanalysisdemonstratedthelysinecontentoftheTOgenerationtransgeniclettucehasbeendifferentlyimprovedandinoneofthemostincreasedby95％GeneticanalysisforsomeoftheT1

10、generationtransgenicplantsbyPCRdisplayedthatthetargetgenecaninheritintoT1generationtransgenicplantsButtherateofgeneticoflysinerichproteingeneinthelettucewaslowKeywards：lysinerichproteingene；transgeniclettuce；lysineconten