金絲桃素合成酶的基因克隆、表達及其酶學性質(zhì)研究.pdf

1、Y774398分類號：密級：單位代碼：10019學號：S02767飛嚼差專夭普碩士學位論文金絲桃素合成酶的基因克隆、表達及其酶學性質(zhì)研究StudyonGeneCloning，ExpressionandPropertyofHypericinSynthetase研究生：王左指導教師：登疊整數(shù)援合作教師：——～一～申請學位門類級別：王堂亟專業(yè)名稱：食昌抖堂皇王猩研究方向：金星生物拉苤所在學院：壘l顯!亞堂皇萱益工程堂瞳AbstractHypc

2、ricinisthemostactivecomponentinHypericumperforatumLIthasthefunctionofantidepression，anti—Vim8’antiturnoutandetcPresentlyhypcricinisproducedbyextractedfromHperforatumI^whichmaycausethelackofwildplantmsoure：eof／／perforatum

3、LTheextractcontainssmallamountofhypericin，mostofitamothercompoundsThesecompoundsmayaccountforsomeofthereportedsideeffectsHypericinsynthetase(Hyp)isanenzymewhichcancatalyzeemodintohypericininacomplexwayIt’sconvenienttopro

4、ducehypcricinbytheenzymeThemainresultwasasfollows：(1)TotalRNAof／／perforatumLwasextractedbyTriz01HypgenewasamplifiedbyRTPCRandclonedintothevectfffofpEI30a，therecombinantpiasmidpET30aHypWastransferredintoEcoliBL210)r巳3)The

5、recombinantstraincouldexpressHypwhenitwasinducedbyIPTGResearchshowedthatHyphadtheabilityofconvertingemodintohypeficin(2)ThefermentativeconditionoftherecombinantstrainwasstudiedThesuitableconditionofshockingbattleofthestr

6、ainwasasfollowsThe州nmrypHofmediumWas7o，theamountofinoculationwasl％andthestrainWasculturedat37“(2for3h，thenH”Wasinducedtoexpressby0。5mMIPTGat30“C、120r／rainfor8h(3)HypWaspurifiedaftercellcollection，suspension，ammoniumsulfa

7、teprecipitationandNiNTAcolumnchromatographyTheenzymeexhibitedhigheractivitywhentemperatureWaS30“CandpHWas7oMchihaelisconstantsof口lzymewere10=5049／Mmol／LVm=O0831m／h‘nag。Keywords：Hypeficinsynthetase，hypericin，Hypericuntper