2023年全國(guó)碩士研究生考試考研英語(yǔ)一試題真題（含答案詳解+作文范文）_第1頁(yè)
已閱讀1頁(yè)，還剩47頁(yè)未讀， 繼續(xù)免費(fèi)閱讀

下載本文檔

版權(quán)說(shuō)明：本文檔由用戶提供并上傳，收益歸屬內(nèi)容提供方，若內(nèi)容存在侵權(quán)，請(qǐng)進(jìn)行舉報(bào)或認(rèn)領(lǐng)

文檔簡(jiǎn)介

1、江南大學(xué)碩士學(xué)位論文PseudomonasputidaS1海藻糖合成酶基因在大腸桿菌中的表達(dá)及其條件研究姓名：姚林申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：生物化工指導(dǎo)教師：毛忠貴段作營(yíng)20080601AbstractAbstractTrehaloseisakindofnon—redoingdisaccharideandiswidespreadinnatureBecausetrehalosehasspecialphysical，chemicalprope

2、rtiesandbiologicalfunction，itcouldbewidelyusedinmolecularbiology，food，cosmetic，pharmaceuticalindustryandagricultureetcTrehalosesynthaseCallconvertmaltoseintotrehalosebyintramoleculartransglucosylationItisoneofthemostimpo

3、rtantwaysaboutthesynthesisoftrehaloseT論SgeneencodingtrehalosesynthaseweresuccessfullyamplifiedfromPseudomonasputidaS1byPeR111eamplifiedfragmentwerelinkedtopMD19TvectorbyTAcloningmethod，andthentransformedthecompetentcells

4、EcoliJM109Thetmnsformantswerescreenedonplatewhichhaveamplicillin／Xgal—IPTGandgettherecombinantplasmidpMDTS2Afteridentifyingtherecombinantplasmidbyrestrictionenzymedigestanalysisandsequencing，usingrestrictionendonucleases

5、BamHIand月砌dIIIdigestbotllpMDTS2andPQE30TObjectgenefragmentwererecollectedandlinkedtoeachotherbyT4DNAligaseIdentifiedbyenzymedigest，therecombinantplasmidpQE—TS2wereconstructed111ecompetentcellsEcoliM15weretransformedwithp

6、lasmidpQETS2andinducedbyIPTGDetectedbySDSPAGE，thetrehalosesynthasegenewasexpressedinEcolianditsmolecularweightwereabout77kDabutthemwereexistedintheformofinclusionbodyinthehosteellsTheconditionsforEcoliM15(pQETS2)cellgrow

7、thandexpressioninshakeflaskwereoptimizedTheoptimizedconditionisthatwhenthecellsgrewintoOD600≈06，IPTGwasaddedtothefinalconcentrationof001mmol／Landlastfor20hoursat20℃Theexpressionoftrehalosesynthasecouldbeashighas186％ofthe

8、totalsolubleproteinsTheactivityoftrehalosesynthaseinthefermentationbrothwasdetectedas19U／mLwhichwasabout50foldhigherthanthatoforiginalstrainnleoptimizedmediumsuitableforhighdensityfermentationiscomposedofglucose10g／L，(NH

9、4)28043g／L，peptone15∥L，citricacid159／L，KH2P041259／L，MgS04‘7H20lg／L，traceelementssolution(TES)5mL／LTheeffectofdissolvedoxygen(DO)concentrationandthefeedingofcarbonandnitrogensourceforthegrowthofEcoliM15(pQETS2)werestudied

10、ata5LfermentorWhentheDOwascontrolledabove30％，carbonandnitrogensourcewasfeededatlogphase，thedrycellweightreached172egLatlastTheexpressionleveloftrehalosesynthasereached102％ofthetotalsolubleproteinwhichwas69timesthanthat、Ⅳ

11、itllflaskfermentationinunitvolumeRecombinanttrehalosesynthasecouldconvertsubstratemaltoseintotrehaloseinonestepTheinfluencesofbufferpH，temperature，reactiontime，substrateconcentration，theamountofenzymeontheconversionofthe

12、maltosewerestudiedItsoptimumpHandoptimumtemperaturewerepH85and20“C，respectivelyTheenzymewasstableunder30℃andbetweenpH75topH90ItshouldbestoredatlowtemperaturesThereactingconditionsoftrehalosesynthasewereresearchedthrought

13、heorthogonaltest。neresultsshowedthat：theoptimumconditionsforconversionofmaltosebytrehalosesynthasewere：pH85，temperature10℃，theamountofenzyme6U／gmaltose，thelastingtimeofreaction36hoursUndertheseconditionsthedegreeofconver

溫馨提示

  • 1. 本站所有資源如無(wú)特殊說(shuō)明，都需要本地電腦安裝OFFICE2007和PDF閱讀器。圖紙軟件為CAD,CAXA,PROE,UG,SolidWorks等.壓縮文件請(qǐng)下載最新的WinRAR軟件解壓。
  • 2. 本站的文檔不包含任何第三方提供的附件圖紙等，如果需要附件，請(qǐng)聯(lián)系上傳者。文件的所有權(quán)益歸上傳用戶所有。
  • 3. 本站RAR壓縮包中若帶圖紙，網(wǎng)頁(yè)內(nèi)容里面會(huì)有圖紙預(yù)覽，若沒(méi)有圖紙預(yù)覽就沒(méi)有圖紙。
  • 4. 未經(jīng)權(quán)益所有人同意不得將文件中的內(nèi)容挪作商業(yè)或盈利用途。
  • 5. 眾賞文庫(kù)僅提供信息存儲(chǔ)空間，僅對(duì)用戶上傳內(nèi)容的表現(xiàn)方式做保護(hù)處理，對(duì)用戶上傳分享的文檔內(nèi)容本身不做任何修改或編輯，并不能對(duì)任何下載內(nèi)容負(fù)責(zé)。
  • 6. 下載文件中如有侵權(quán)或不適當(dāng)內(nèi)容，請(qǐng)與我們聯(lián)系，我們立即糾正。
  • 7. 本站不保證下載資源的準(zhǔn)確性、安全性和完整性, 同時(shí)也不承擔(dān)用戶因使用這些下載資源對(duì)自己和他人造成任何形式的傷害或損失。

評(píng)論

0/150

提交評(píng)論