重組克隆技術中植物啟動子分析載體的改進研究.pdf

1、浙江大學碩士學位論文重組克隆技術中植物啟動子分析載體的改進研究姓名：金煒元申請學位級別：碩士專業(yè)：遺傳學指導教師：王君暉20070611浙江大學碩上學位論文ABSTRACTPromoterisakindofDNAregionorsequencethatcancombinewithRNApolymeraseandothertranscriptionfactorsThepromoteranalysisisaveryimportantande

2、fficientmethodusedingenesfunctionanalysisandspecificexpressionresearchFirstweconstructed17promoteranalysisvectorsof12differentgenepromotersusingthetraditionalcloningsystembasedOilrestrictionfinzymesandligaseThoughmostoft

3、heconstructionsse躑ltogetsuccessfulresultswestillfoundlotsofexistentandpotentialproblemsthroughdeepanalysisofthesesequencesandtransgenicmaterialsGusstainingresultsThe5’untranslafionregionsegmentlengthespeciallytheTATAboxa

4、ndtheputativeATGbetweenthenativeATGandthetranscriptionstartsitehaveimportantinfluencesonthefinalsucaT／鼯$The2x35SpromoteroftheresistantganeupstreamalsointerfereOUrexperimentreliabilityBesideswealsofoundthatthetransgenicli

5、nesweresodistinctivetoeachotherthatitishardforllstogetanuniqueandreliableresultNowwehavetransferredtoanewcloningsystemcalledGatewayTechnology衄inordertogetbetterefficientpromoteranalysisvectorsWhereasthefactknownsOfarthat

6、thetransgenvariationmainlyraisedbythedistinctchromatinenvironmentsandvariousactivateorinhibitsignalsespeciallythehcterochromatinsurroundingthedifferentinsertionsitesⅥ，llileinsulaterisakindofDNAbarrierelementthatpossesses

7、acornsionabilitytoprotectgenesfrominappropriatesignalsemanatingfromtheirsurroundingenvironmentSoitiswidelyusedintransgensinordertominimizethetransgenievariationInfactithasbeenusedsuccessfullyandwidelyintransgenicmoose，ra

8、bbitandcelllinesHoweverbecauseoftbelaterbeginninginplantinsulatorresearchithasfewreportsabouttheapplicationsofinsulatorsinplantnansgensInthispaperwedidsuchapathbreakingworkbyaddingadrosophilainsulatorsequencetoourGateway