含有MAR序列的STI-STII-LTB融合基因在苜蓿中的表達(dá).pdf

1、分糞號密級石河子大學(xué)碩士學(xué)位論文y一9圣185zt含有MAR序列的STISTIILTB融合基因在苜蓿中的表達(dá)研究生武冬梅陳創(chuàng)夫教授指導(dǎo)教精——祝建波副教授申請學(xué)位門類級別理學(xué)碩士學(xué)科、專業(yè)名稱生物化學(xué)與分子生物學(xué)研究方向植物基因工程技術(shù)應(yīng)用所在學(xué)院生命科學(xué)學(xué)院中國新疆t石河子2006年5月AbstractInthisreport，firststep，MARswereobtainedfromthegenomeoftobaccobyPCRB

2、glucuronidase(aug)geneofplantexpressionvectorpBll21wasflankedbytheMARtoformanewplantexpressionvectorSTI，STIIgenewereobtainedfromplasmidpGEM5Zf()STIandK88ab(L1阡，ST十)Theheat—stabletoxin(STlSTII)werefusedbyTP—PCR，andthenfus

3、edheat—labiletoxinBsubunitbyrestrictionendonucleaseThe5‘terminusofthegeneencodingLTBwasfusedtothe3’terminusoftheSTgene，asevenaminoacidlinkerwereinsertedbetweentheSTmadLTBgeneThefusedgeneSTISTIILTBwasclonedintoplantexpres

4、sionvectorpBll21一MARsteplaceGUSgene，anotherplantexpressionvectorwasconstructedwithoutMARsequenceRecombinantplasmidswereamplifiedbyPCR，restrictionendonucleaseanalysisandDANsequencingTheresultsindicatethattherecombinantpla

5、smidswerecorrectlyconstructedTherecombinantplasmidspBll21MARs—STISTIILTBpBll21STISTILLTBweretransferredintoAgrobacteriumtumefaciensEHAl05ByusingCO—cultivatedmethod，thefusedgeneSTI—STII—LTBandMARsweretransferredintoalfalf

6、acellTransferredshootswereselectedOUsolidmediumcontainingKanamycinTransgenicalfalfaplantswerefoundcontainingSTISTIILTBgenebyPCRandSouthernblottingThesetransgenicalfalfaplants，wereconfumedexpressingSTISTHLTBrecombinantpro

7、teinthroughSDS—PAGEBasedonWesterndotting，resultsshowedthatrecombinantproteinexpressedbytransgenicalfalfaplantscouldelicitingimmunoresponsetoLTBantiboayThisprojectcouldoffersciencewarrantforstudyingediblevaccineKeywords：m