波爾山羊體細(xì)胞核移植研究.pdf

1、分類號密級單位代碼：!Q3鯉學(xué)號：墨業(yè)Q9Q5Q1QQ2瘩微雇業(yè)大學(xué)碩士學(xué)位論文波爾山羊體細(xì)胞核移植研究StudyonSomaticCellNuclearTransferinBoerGoat研究生：韓偉指導(dǎo)教師：章孝榮教授合作指導(dǎo)教師：圍耍副數(shù)援申請學(xué)位門類級別：專業(yè)名稱：研究方向：所在學(xué)院：盔堂亟墊塑塑堡直登皇鳘墮塾塑墅墮三猩動物科技學(xué)院答辯委員會主席二Oo六年六月Abstractnlepresentstudywasconducted

2、toinvestigatethesomaticcellnucleartransferinBoergoat，such懿donorcellculture，recipientoocyteinvitromaturation，reconstructedooeyteelectrofusionandactivation，embryoinvitroculture，andembryotransferTheexperimentsuccessfullypri

3、maryculturedandsubculturedearskinfibroblastsdedvedfromaBoergoatbytissueculturenledifferentfreezingprotocolswereinvestigatedtocryopreservethefibroblastsFirst，thefibroblastcellswereequilibratedat4“Cforlh，andstoredat20“Cfor

4、6h，andthenplungedliquidnitrogenSecond，thecellswerewereequilibratedat4“Cforl垃storedat20“(2for6hand80℃overnight，andthenplungedintoliquidnitrogenThird，thecellswereequilibratedat4“Cforlh，keptat80“Covernightbeforebeingplunged

5、intoliquidnitrogenTheresultshowedthattheadherenceratesweremostsignificantlydifferentbetweenthemafterthawingandculture(238％，2281and5733％respectivelyPO05)TheextrusionrateofPBlofgoatoocytesWaSsignificantlyhigherwhentheOOCyt

6、eswe他cultured21—25hthanthatof17h(P005；870％VS750％，P005)Whentheclonedembryoswerecoculturedwithfetalmousefibroblastmonolayertheblastocystdevelopmentwas870％butnoblastocystformationwereobtainedwithoutCOcultureThereconstructed

7、embryoswereculturedfor13h，36h，69hrespectivelyaRcrfusion。andthenactivationwasundertakenbyusingionomycinand6DMAPWefoundthatthecleavageratehadnosignificantdifferencebetween13hgroupand36hgroup(7258％VS7297％PO05)，butboththerat