1、北京化工大學碩士學位論文釀酒酵母S腺苷甲硫氨酸合成酶基因的克隆及表達研究姓名：安勝欣申請學位級別：碩士專業(yè)：生物化工指導教師：王雅琴20060612北京化r大學碩士學位論文CloningandexpressionofS—adenosylmethioninesynthetase“n■●‘genefrom3nccnQromvcescerevtstaeABSTRACTSadenosylmethionine(SAM)iSanimportantm

2、etaboliteinvivoandtakespartinmorethanfortykindofbiochemicalreactionssuchasthegeneexpression，cellmembranefluidityandthesynthesisofpeptideClinicalstudiesshowedthatSAMhascurativeeffectsOilosteoarthritis，depressionandthediso

3、rderofliverfunctionSAMhasalsobeenfoundtobeanadvancedhealthcareproductwhichcanpreventcancer，cardiovasculardiseasesandanti—senilityTherearemanypatientswithhepatitisinourcountry，andthenumberofpatientswithdepressionorosteoaa

4、hritisisincreasingalongwiththefasterlivingrhythmSOSAMhasabroadermarketButwehavenotyelbeenabletoindustrialproductioninChinaThou曲thereareenormousSAMcommoditiestosupplyforthepeopleneeding，itcannotbepopularizedbecauseofitsex

5、pensivepriceThusitisurgetoexploreahighproducingandlowcostproducingtechnologyThegeneencodingsam2wasamplifiedfromSaccharomycescerevisiae2B41chromosomalDNAusingPCRtechniqueonthebaseofthespecificprimersfromSaccharomycescerev

6、isiaeinGenBankThePCRproductwaftapproximately1200bpDNAsegmentUsingTAcloningtechnique，thePCRproductwasclonedintothepUCmTvectorandthecloningplasmidpUCmTsam2wasconstructedsuccessfullypUCm—Tsam2andpET28a(]weredigestedbyEcoRIa

7、ndHindIIIdoubleenzymesThepurifiedSAM2genewasclonedintotheexpressionvectorpET28a()，andtheprokaryoticexpressionvectorpETsam2wasthusconstmctedTherecombinantplasmidpETsam2wastransformedintocompetenceEcoliBL2l(DE3)strainTheba