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《茶樹脫水素基因dhn2的原核表達和體外功能鑒定》由會員上傳分享，免費在線閱讀，更多相關(guān)內(nèi)容在學術(shù)論文-天天文庫。
1、AbstractDehydrinwasfirstlyfoundin1980s．whichisakindoflateembryogenesisabundantproteins．ItbelongstothefamilyofLEAD-IIanditsmolecularweightchangingfrom9to200KD．Inrecentyears．theresearchofdehydrinadvancedrapidly,espciallytheirprotectingfunctiontotheplantsattractedmoreandmorepepole’Sattention．T
2、hedehydrinisabundantinglycinandlysine，becauseofthatdehydrinsarehighlyhydrophilicandCancombinewithcellmembraneeasilyinordertopreventexcessivelywatererosion．ThatisthereasonwhydehydrinCanprotectcellsfromthedamageofdrought．Inaddition．proteinscanalsobeenprotectedbydehydrinsbecauseoftheirdoubleaf
3、finitystructureofalphahelixservingseemlyasmolecularchaperone．CamelliasinensisisakindofPerennialeconomiccropsinsubtropicalareaandvulnerabletocold．Sohowtoenhancetheteaplant’Sabilityofanti-coldhasattractedmoreandmoreattention．ThefragmentofonedehydrinwasscreenedformSSHlibraryofteainducedbycold．
4、AfteramplifiedbyRACEkitandsequenced．thedehydnnwasidentifiedasSK2typeandthennameddhn2．Theresultsofthestudiesarelistedasfollows：Firstly,thetranscriptionsofdhn2underthestressesofcold，dehydrationandABAweretestedbythemethodofRT-PCR．Theresultshowedthatdhn2couldbeinducedbycold，dehydrationandABA，in
5、whichtheinducedeffectofcoldanddehydrationwereveryobvious．Hence，itcouldspeculatedthatdhn2playsimportantroleinresistanceofcoldanddrought．Secondly,procaryoticexpressionvectorofdhn2wasconstructedsuccessfully．Afterinducedby0．8mmol／LIPTGat30。C，a53KDcombinantproteinwasobtainedandwaspositivetoantib
6、odyanti—dehydrin．So，theproteincouldbecertifiedasdhn2recombinantprotein．Thepurityofdhn2afterpurificationisaround90％．Finally,invitrotesting，theprotectivefunctionofthedehydrinproteinwasveryobvious，theresidualenzymeactivityofMDHandLDHstillremained86％，62％respectively,afterdehydrationandfreezingi
7、nactivation．ComparedwithBSA，dhn2hadhigherabilityofanti—droughtbutlowerabilityofanti-freeze．Keywords：Camelliasinensis；Dehydrin；Real—timePCR；Prokaryoticexpression；Proteinpurification；Westernblotting；FunctionalcharacterizationinvitroII圖表清單圖1．1脫水素基因表達信號轉(zhuǎn)導示意圖
