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《脂質(zhì)體介導(dǎo)的cho細(xì)胞轉(zhuǎn)染——脂質(zhì)體介導(dǎo)的基因真核細(xì)胞轉(zhuǎn)染 畢業(yè)論文》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(kù)。
1、脂質(zhì)體介導(dǎo)的CHO細(xì)胞轉(zhuǎn)染——脂質(zhì)體介導(dǎo)的基因真核細(xì)胞轉(zhuǎn)染專(zhuān)業(yè)：生物工程專(zhuān)業(yè)班級(jí)：2008級(jí)二班姓名：30摘要CHO細(xì)胞是中國(guó)倉(cāng)鼠卵巢細(xì)胞，是目前生物工程上廣泛使用的細(xì)胞系。真核細(xì)胞中CHO細(xì)胞是目前重組糖基蛋白生產(chǎn)的首選體系。外源基因進(jìn)入細(xì)胞主要有四種方法：電擊法、磷酸鈣法和脂質(zhì)體介導(dǎo)法和病毒介導(dǎo)法。脂質(zhì)體是磷脂分散在水中時(shí)形成的脂質(zhì)雙分子層，又稱(chēng)為人工生物膜。根據(jù)膜的融合及內(nèi)吞作用，可用作外源物質(zhì)進(jìn)入細(xì)胞的載體。在對(duì)生物大分子的傳遞過(guò)程中，脂質(zhì)體是十分有效的載體，無(wú)論對(duì)微生物還是對(duì)植物細(xì)胞或?qū)Σ溉閯?dòng)物細(xì)胞導(dǎo)入外源DNA，脂質(zhì)體的包裹都能對(duì)抗核酸酶的消化。脂質(zhì)體的制備主要有
2、超聲波法、磷脂酰絲氨酸Ca2+離子融合法和逆相蒸發(fā)等，用超聲波法可制備的單層或多層脂質(zhì)小體能向細(xì)胞內(nèi)引入的物質(zhì)量少，故常用后兩種方法制備大的脂質(zhì)體。脂質(zhì)體介導(dǎo)轉(zhuǎn)染法的改進(jìn)方法包括促進(jìn)轉(zhuǎn)染的脂類(lèi)、增加轉(zhuǎn)染效率的添加劑、DNA與脂質(zhì)體最佳比率的確定。本實(shí)驗(yàn)將攜帶綿羊P基因的綠色熒光蛋白真核表達(dá)載體轉(zhuǎn)入中國(guó)倉(cāng)鼠卵巢細(xì)胞，得到穩(wěn)定表達(dá)綿羊PrP基因的CHO細(xì)胞系。得出外源導(dǎo)入CHO的綿羊PrP可以在CHO中穩(wěn)定表達(dá)的結(jié)論。關(guān)鍵詞：CHO細(xì)胞、脂質(zhì)體、轉(zhuǎn)染30AbstractCHOcellsofChinesehamsterovarycells,biologicalengineering
3、iscurrentlywidelyusedcelllines.EukaryoticcellsinCHOcellsiscurrentlyrestructuringglycosylproteinofchoicefortheproductionsystem.Exogenousgenesintocellsmainlyhasfourkindsofmethods:electricshockmethod,calciumphosphateandmediatedbyliposomeandvirusmediatedmethod.Liposomeisaphospholipiddispersedin
4、waterisformedwhenthelipidbilayer,alsoknownastheartificialbiologicalmembrane.Accordingtothemembranefusionandendocytosis,canbeusedasexogenoussubstancesintothecellcarrier.Onbiologicalmacromoleculesinthetransferprocess,liposomesareveryeffectivecarrier,regardlessofthemicroorganismorplantcellorma
5、mmaliancellsforintroducingexogenousDNA,liposomeencapsulatedcanagainstnucleasedigestion.Preparationofliposomesareultrasonicmethod,phosphatidylserineCa2+ionfusionmethodandreversephaseevaporation,usingultrasonicmethodtopreparemonolayerormultilayerlipidglobulestointracellularintroducedmassisles
6、s,socommonlyusedtwomethodsforpreparationoflargeliposomes.Liposomemediatedtransfectionmethodimprovementmethodsincludepromotingtransfectionlipids,increaseintransfectionefficiencyadditives,DNAandliposomebestratedetermine.ThisexperimentwillcarrysheepPgenegreenfluorescentproteineukaryoticexpress
7、ionvectorwastransfectedintoChinesehamsterovarycells,stableexpressionofPrPgeneinsheepintheCHOcellline.DrawforeignimportCHOsheepPrPcanCHOstableexpressionoftheconclusion.Keywords:CHOcells,liposomes,transfection分享到翻譯結(jié)果重試抱歉，系統(tǒng)響應(yīng)超時(shí)，請(qǐng)稍后再試·支持中英、中日在線互譯·支持網(wǎng)頁(yè)翻譯，在輸入
