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《茶樹脫水素基因dhn2的原核表達(dá)和體外功能鑒定(6216)》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、AbstractDehydrinwasfirstlyfoundin1980s．whichisakindoflateembryogenesisabundantproteins．ItbelongstothefamilyofLEAD-IIanditsmolecularweightchangingfrom9to200KD．Inrecentyears．theresearchofdehydrinadvancedrapidly,espciallytheirprotectingfunctiontotheplantsattractedmoreandmorepepole’Sattention．Thedehydr
2、inisabundantinglycinandlysine，becauseofthatdehydrinsarehighlyhydrophilicandCancombinewithcellmembraneeasilyinordertopreventexcessivelywatererosion．ThatisthereasonwhydehydrinCanprotectcellsfromthedamageofdrought．Inaddition．proteinscanalsobeenprotectedbydehydrinsbecauseoftheirdoubleaffinitystructureo
3、falphahelixservingseemlyasmolecularchaperone．CamelliasinensisisakindofPerennialeconomiccropsinsubtropicalareaandvulnerabletocold．Sohowtoenhancetheteaplant’Sabilityofanti-coldhasattractedmoreandmoreattention．ThefragmentofonedehydrinwasscreenedformSSHlibraryofteainducedbycold．AfteramplifiedbyRACEkita
4、ndsequenced．thedehydnnwasidentifiedasSK2typeandthennameddhn2．Theresultsofthestudiesarelistedasfollows：Firstly,thetranscriptionsofdhn2underthestressesofcold，dehydrationandABAweretestedbythemethodofRT-PCR．Theresultshowedthatdhn2couldbeinducedbycold，dehydrationandABA，inwhichtheinducedeffectofcoldandde
5、hydrationwereveryobvious．Hence，itcouldspeculatedthatdhn2playsimportantroleinresistanceofcoldanddrought．Secondly,procaryoticexpressionvectorofdhn2wasconstructedsuccessfully．Afterinducedby0．8mmol／LIPTGat30。C，a53KDcombinantproteinwasobtainedandwaspositivetoantibodyanti—dehydrin．So，theproteincouldbecer
6、tifiedasdhn2recombinantprotein．Thepurityofdhn2afterpurificationisaround90％．Finally,invitrotesting，theprotectivefunctionofthedehydrinproteinwasveryobvious，theresidualenzymeactivityofMDHandLDHstillremained86％，62％respectively,afterdehydrationandfreezinginactivation．ComparedwithBSA，dhn2hadhigherability
7、ofanti—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(dá)信號(hào)轉(zhuǎn)導(dǎo)示意圖
