重組克隆技術(shù)中植物啟動子分析載體的改進(jìn)研究.pdf

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