波爾山羊體細胞核移植研究.pdf

1、分類號密級單位代碼：!Q3鯉學號：墨業(yè)Q9Q5Q1QQ2瘩微雇業(yè)大學碩士學位論文波爾山羊體細胞核移植研究StudyonSomaticCellNuclearTransferinBoerGoat研究生：韓偉指導教師：章孝榮教授合作指導教師：圍耍副數(shù)援申請學位門類級別：專業(yè)名稱：研究方向：所在學院：盔堂亟墊塑塑堡直登皇鳘墮塾塑墅墮三猩動物科技學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