簡介：上海水產(chǎn)大學(xué)碩士學(xué)位論文大黃魚腎組織細(xì)胞系PCK的建立及其生物學(xué)特性姓名陳少鵬申請學(xué)位級(jí)別碩士專業(yè)水產(chǎn)養(yǎng)殖指導(dǎo)教師楊先樂20040420ESTABLISHMENTANDCHARACTERIZATIONOFACEILLINEFROMABSTRACTTHISPAPEL’NASABOUTHOUTOESTABLISHACELLSTLALN1INE0LLPVCLTDO．SCITLTTLI“Y，“，C，THROUGHPRIMARCULTM’EANDTOSTUD＼ITSCHARACTERIZATIONINORDERTODRO～I(xiàn)DERELIABLEMETHODSTOSOLVETHEPL’OBLEMI11THERESEARCHANDMANUFACTURETHEHEALTH＼PSELTDOSTIAENAC‘J1，LELIABOUTTV,OVEARSOLDWEREADOPTED．ANDTHEMETHODSOFZOU、ENGONGWEREREFERREDTHECELLSOFPRIMARYCULTUREWEREOBTAINEDSUCCESSFULL、FFOMTHEPROBOSCIS．KIDNEY．1I、_ERANDDORSALFINOFPSEUDOXCIAEML【’Ⅲ【‘C“THECELILINEFROMTHEKIDNEYOFTHECEILSOFOFNCLIDOSCIAETQACROCEAWASESTABIISHEDANDNAMEDPCKCELLLINE．PCKCELLLINEWASCHARACTERIZED．PCKCELLLINEISFIBROBLAST．RHECELLSWERESUBCULTUREDINM199MEDIUMSUPPLEMENTEDWITH5％一10％NBS．THEOPTIMUMTEMPERATUREXVAS2426℃．ITSPHRANGEDFROM65T07．2THEPOPULATIONDOUBLETIMEWAS557H．THEAVERAGESUBCULTURETIMEWAS3D，THECONDITION1{PRESERVATIONWASSTUDIEDITWASINDICTEDTHATPCKCELLLIFIEWASN’TFITFORPRESERVEDAT15℃AND4℃．BUTITCOULDPRESERVEDINA習(xí)0‘℃舶EZERANDINLIQUIDNI仃09ENCONTAINER．7西PPROCESSOFPRESEI‘‘ATIONINLIQUIDNITROGENCONTAINERH孫BALANCINGLHAT4。C．FREEZINGAL70。CANDPUTTINGINTOLIQUIDNITROGEN．111EKARYOLOGYOFPCKWASANALYZED．ITWASSHOWEDTHATTHEADIPLOIDNTTMBERRMAGINGFROM31TO55V／ITHAMODALPEAKAT48CHROMOSOMESATPASSAGEL1．THECHROMOSOMENUMBERDISTRIBUTIONATPASSAGE63DISPLAYEDATA2NVALUERANGINGBETWEEN32TO86．ANDTHEMETAPHASESWITHTHENORRNALDIPLOIDNUMBERWASABOUT14％NEMETHODSOFPRIMARYCULTURETHECELLSFROMTHELIVERCOULDOBTAININTHEDIGESTIONMETHODS．THOSEFROMTHEPROBOSCISANDTHEFINSCOULDOBTAININTHETISSUEMETHODS．ANDTHOSETBAMTHEKIDNEYCOULDOBTAININBOTHMETHODS．SO．THECELLSFROMDIFFERENTTISSUESSEEMEDBE倚T’ORTHEDIFFERENTPRHARYMETHODS．THEEFFECTOFSCLUMONTHECELLS111ECELLSHADLOWDEMANDONTHERATESANDCONTAINSOFSERUM．THEMEDIUMWITH20‰NBSCOULDBEUSEDINDRIMANCULTURE．ANDWHICHWITH5％NBSWEREUSEDINCULTUREPCKCELLLINECHARACTERIZATIONOFPCKCELLLINEPCKCELLSWEREEPITHELIUM．BIASLANDFIBROBLASTINTHEPRIMMCULTURE．WITHSUBCULTUMTHE3MADUALLYBECAMETHEFIBROBLASTCELLS．THECELLSGREVERYFAST．GENERNLY．ITCOULDFORMTHEMONOLAYERIN3D．KARYOTYPEREVEALEDTHATPCKCELLLINEEREANANEUPLOIDCELLLINCKEYWORDSPSETTDO．SCIAETLCICN’CEU．PRIMADCTLLTTLRELPCK．DOUBLINGLIMECHROMOSOLNECR,,OPRESERVATION

簡介：學(xué)位論文獨(dú)創(chuàng)性聲明本人鄭重聲明1、堅(jiān)持以“求實(shí)、創(chuàng)新”的科學(xué)精神從事研究工作。2、本論文是我個(gè)人在導(dǎo)師指導(dǎo)下進(jìn)行的研究工作和取得的研究成果。3、本論文中除引文外，所有實(shí)驗(yàn)、數(shù)據(jù)和有關(guān)材料均是真實(shí)的。4、本論文中除引文和致謝的內(nèi)容外，不包含其他人或其它機(jī)構(gòu)己經(jīng)發(fā)表或撰寫過的研究成果。5、其他同志對本研究所做的貢獻(xiàn)均已在論文中作了聲明并表示了謝意。作者簽名日期等耘2ALLO豐5TO學(xué)位論文使用授權(quán)聲明本人完全了解南京師范大學(xué)有關(guān)保留、使用學(xué)位論文的規(guī)定，學(xué)校有權(quán)保留學(xué)位論文并向國家主管部門或其指定機(jī)構(gòu)送交論文的電子版和紙質(zhì)版有權(quán)將學(xué)位論文用于非贏利目的的少量復(fù)制并允許論文進(jìn)入學(xué)校圖書館被查閱有權(quán)將學(xué)位論文的內(nèi)容編入有關(guān)數(shù)據(jù)庫進(jìn)行檢索有權(quán)將學(xué)位論文的標(biāo)題和摘要匯編出版。保密的學(xué)位論文在解密后適用本規(guī)定。作者簽名摘要綜上所述，與親本“小堰54”和“8602”相比，雜交小麥新品種“小堰81的莖稈的高度、解剖結(jié)構(gòu)和細(xì)胞壁的木質(zhì)素與纖維的含量和組成等莖稈的質(zhì)量都更有利于抵抗風(fēng)雨等外力的作用，從而減少倒伏的發(fā)生。對高產(chǎn)小麥品種“京411”的旗葉和芒的葉綠體發(fā)育過程的研究表明芒對光合作用的貢獻(xiàn)也不容忽視。關(guān)鍵詞小麥莖稈抗倒伏解剖木質(zhì)素纖維葉綠體

簡介：西北農(nóng)林科技大學(xué)碩士學(xué)位論文耳廓軟骨細(xì)胞生物學(xué)特性研究及組織工程軟骨的體外構(gòu)建初探姓名溫葉飛申請學(xué)位級(jí)別碩士專業(yè)臨床獸醫(yī)學(xué)指導(dǎo)教師張涌200406012與未完全消化的組織塊共培養(yǎng)可為軟骨細(xì)胞的生長提供一個(gè)與正常生理?xiàng)l件相似的生長環(huán)境，有利于軟骨細(xì)胞經(jīng)多次傳代后的正常增殖和表型維持。3，用BFF替代FBS培養(yǎng)軟骨細(xì)胞能較好地促進(jìn)細(xì)胞增殖，并能維持軟骨細(xì)胞的表型，以添加L0?F的培養(yǎng)液促增殖效果最好。4添加EGF也能提高軟骨細(xì)胞的增殖速率，但對軟骨細(xì)胞去分化現(xiàn)象沒有明顯的響，即不能抑制軟骨細(xì)胞向成纖維樣細(xì)胞的改變。5在離心管中無支架培養(yǎng)得到外觀呈軟骨樣的組織塊。關(guān)鍵詞軟骨細(xì)胞牛卵泡液表皮生長因子組織工程軟骨。

簡介：耗牛周期黃體退化過程中黃體細(xì)胞凋亡的生物學(xué)特征摘要本研究采用不同的手段和研究方法，通過對耗牛周期黃體退化過程中組織形態(tài)學(xué)的觀察、體外培養(yǎng)條件下周期黃體細(xì)胞凋亡的生物學(xué)特征的分析、TNFA對體外培養(yǎng)周期黃體細(xì)胞凋亡的影響以及周期黃體退化過程中黃體細(xì)胞內(nèi)CMYC和BAD的表達(dá)的測定等，系統(tǒng)地研究了耗牛周期黃體退化過過程中細(xì)胞凋亡的生物學(xué)特征，取得了如下結(jié)果1采集不同階段耗牛的周期黃體，進(jìn)行13E染色、光鏡觀察、超微結(jié)構(gòu)觀察和DNA原位熒光標(biāo)記檢測DAPI，發(fā)現(xiàn)整個(gè)周期黃體期均有黃體細(xì)胞凋亡現(xiàn)象，但主要見于發(fā)情期黃體。隨著黃體老化，黃體組織中的血管也發(fā)生退化。2用酶消化法制備耗牛原代黃體細(xì)胞，經(jīng)血清撤除誘導(dǎo)其凋亡，然后檢測其生物學(xué)特征。采用原位熒光標(biāo)記DAPI和PI法法檢測發(fā)現(xiàn)，凋亡細(xì)胞主要表現(xiàn)為核固縮、并發(fā)散高亮的藍(lán)色熒光DNA原位末端標(biāo)記檢測表明，凋亡細(xì)胞呈TUNEL陽性反應(yīng)透射電鏡觀察顯示，凋亡細(xì)胞核固縮、染色質(zhì)邊集、核碎裂和凋亡小體形成DNA電泳呈現(xiàn)典型的“梯狀帶”，與己知分子量的MARK對比，證實(shí)凋亡的黃體細(xì)胞中存在分子量為180200BP整倍數(shù)的DNA片段。3在培養(yǎng)液中，添加不同劑量的TNFA，均能誘導(dǎo)體外培養(yǎng)的耗牛周期黃體細(xì)胞發(fā)生凋亡，并呈劑量依賴性流式細(xì)胞術(shù)FCM檢測，可見明顯的凋亡峰。4用免疫組化法檢測了周期黃體不同階段CMYC和BAD的表達(dá)情況，發(fā)現(xiàn)在整個(gè)周期黃體期，黃體細(xì)胞中OMY。和BAD均有不同程度的表達(dá)，并且均定位于細(xì)胞漿中。在周期黃體存活的過程中，隨著其存活時(shí)間的延長，黃體細(xì)胞的陽性表達(dá)率和表達(dá)強(qiáng)度也呈逐漸增大的趨勢，并在發(fā)情前期和發(fā)情期黃體達(dá)到相對穩(wěn)定的表達(dá)狀態(tài)在發(fā)情期黃體中，部分黃體細(xì)胞的核內(nèi)也有CMYC蛋白存在。以上結(jié)果表明，黃體細(xì)胞凋亡是導(dǎo)致耗牛周期黃體退化的主要原因耗牛黃體細(xì)胞可以在體外培養(yǎng)血清撤除能誘導(dǎo)其凋亡凋亡的黃體細(xì)胞在形態(tài)學(xué)和生化等方面具有典型的特征TNFA能誘導(dǎo)體外培養(yǎng)的耗牛黃體細(xì)胞發(fā)生凋亡，并呈劑量依賴性CMYC和BAD基因的表達(dá)可能參與耗牛黃體細(xì)胞凋亡的分子調(diào)控。關(guān)健詞耗牛，黃體細(xì)胞，細(xì)胞凋亡，體外培養(yǎng)，CMYCBADTNFATUNEL原位熒光標(biāo)記4IMMUNOHISTOCHEMISTRYWASUSEDTODETECTTHEEXPRESSIONOFCMYCANDBADOFTHECYCLICCORPUSLUTUMINDIFERENTSTAGESIMMUNOSTAININGWASSEENINCYTOPLASMOFLUTEINCELLSINWHOLELUTEALPHASEFURTHERMORETHEPOSITIVERATEANDIMMUNOSTAININGINTENSITYINCREASEDGRADUALLYWITHTHEDEVOLOPMENTOFCORPUSLUTEURNTHELEVELSOFEXPRESSIONWERERELATIVELYSTABLEINESTROUSSTAGECMYCPROTEINWASALSOFOUNDINTHENUCLEIOFLIMITEDNUMBEROFLUTEALCELLSITISCOUCLUDEDFROMTHERESULTSMENTIONEDABOVETHATAPOPTOSISOFLUTEALCELLSPLAYSAMAJORROLEINTHEREGRESSIONOFCYCLICCORPUSLUTUMINYAKLUTEALCELLSCANBECULTUREDINVITROANDBEINDUCEDTOAPOPTOSISBYUSEOFSERUMWITHDRAWINGMETHODTNFAOFDIFERENTCONCENTRATIONCANALSOINDUCETHEAPOPTOSISOFLUTEALCELLSINVITROWITHADOSEDEPENDENTMANNERGENEEXPRESSIONOFGMYCANDBADMAYBEINVOLVEDINTHEREGULATIONOFLUTEALCELLSAPOPTOSISKEYWORDSYAKLUTEALCELLSAPOPTOSISINVITROCMYCBADTNFATUNELDAR

簡介：摘要為了闡明馬傳貧驢白細(xì)胞弱毒疫苗DLA．EIAV致弱及免疫保護(hù)的分子機(jī)制，給其它饅病毒的免疫預(yù)防提供借鑒，本實(shí)驗(yàn)室構(gòu)建了DLAEIAV感染性分子克隆命名為POK8266并獲得其衍生病毒命名為VOK8226．同時(shí)對我國馬傳染性貧血病毒強(qiáng)弱毒株進(jìn)行了序列分析。在此基礎(chǔ)上，以POK8226為骨架，構(gòu)建了馬傳染性貧血病毒強(qiáng)塌LTR嵌臺(tái)毒感染性分子克隆命名為POKVLTR，并獲得了其衍生病毒命名為VOKVITR。本研究對VOK8226、VOKVITR和DLA．EIAV進(jìn)行了動(dòng)物接種試驗(yàn)，通過對臨床和病理組織學(xué)變化、各結(jié)構(gòu)蛋白抗體、感染馬體內(nèi)病毒載量的監(jiān)測以及LTR及∞V基因在感染馬體內(nèi)的變異規(guī)律進(jìn)行了研究。將7匹EIAV陰性健康馬分為4組．第1組4和5馬接5MLXL04TCID∞嵌合毒POKVLTR，第2組6和7馬接種5MLXL05TCID∞克隆毒VOK8226，第3組8接種5MLXLONTCID。疫苗毒DLAEIAV，葡時(shí)設(shè)第4組I和2馬作為菲接種對照組。接種后第220天，除了2馬攻毒劑量為3MX104血清稀釋度外，其它所有馬均用3MLXL04血清稀釋度EIAV強(qiáng)毒遼寧株LEIAV進(jìn)行攻擊。攻毒前后對每匹馬進(jìn)行體溫監(jiān)測．發(fā)現(xiàn)在接種后．所有試驗(yàn)馬未見體溫升高現(xiàn)象，說明我們所用的毒株接種后對馬是很安全的，用L．EIAVLTR置換VOK8226的LTR得到的嵌合病毒對馬也沒有表現(xiàn)出臨床致病性攻毒后，非接種對照紐L和2馬體溫均出現(xiàn)典型的稽留熱，并分別于攻毒后第134天和19天死亡。病理剖撿發(fā)現(xiàn)死亡馬呈現(xiàn)典型的馬傳貧的病理組織學(xué)變化。第1組和第3組嵌合毒接種組和疫苗毒接種組在攻毒后未見任何臨床變化，證明嵌合毒和疫苗毒接種馬得到了免疫保護(hù)。第2組克隆毒接種組中的6馬在攻毒后沒有異常變化，而7馬卻經(jīng)歷6次發(fā)熱期，晟終于攻毒后第185天死亡，但開始發(fā)熱和死亡的時(shí)間均比對照組明顯滯后．病理剖檢發(fā)現(xiàn)發(fā)病死亡馬具有典型的馬傳貧的病理組織學(xué)變化。在攻毒后第450天剖殺所有存活馬，并進(jìn)行病理組織學(xué)檢查，結(jié)果表明所有免疫保護(hù)馬均未見任何異常變化。在試驗(yàn)過程中，我們利用ELISA方法對不同試驗(yàn)馬攻毒前后血清中針對EIAV結(jié)構(gòu)蛋白P15、PI1、P26、P9以及GP45的抗體進(jìn)行了檢測。攻毒前各接種馬抗P9、P1】和P15抗體水平極低或檢測不到．P26抗體上升后又逐漸下降。GP45抗體緩慢上升后并持續(xù)在較高水平。攻毒后發(fā)病馬1，2．7體內(nèi)抗P9、P11、P15、P26和GP45抗體均顯著升高，并持續(xù)至死亡，6和8馬體內(nèi)抗P15、P26和GP45抗體也有升高外。攻毒后，嵌合病毒免疫馬4和5體內(nèi)抗這五種結(jié)構(gòu)蛋白抗體和攻毒前沒有鵯顯變化，表明攻毒后沒有回憶反應(yīng)，表明其免疫效果可能優(yōu)于其它各接種組。抗GP45抗體變化在發(fā)病馬和未發(fā)崩馬同無明顯差異．提示GP45抗體變化與疾病進(jìn)程沒有直接的關(guān)系。利用實(shí)時(shí)定量PCR技術(shù)對攻毒前后馬外周血白細(xì)胞中EIAV前病毒DNA及馬血漿中病毒RNA的載量進(jìn)行了監(jiān)測，發(fā)現(xiàn)感染馬體內(nèi)前病毒DNA載量與P15和P26抗體水平有一定的相關(guān)性，血漿中病毒RNA的載黛與P9和PLI抗體水平成正相關(guān)。發(fā)病馬血漿病毒RNA超過106拷貝，MI血漿，而免疫保護(hù)馬血漿中病毒RNA載量較低10”拷貝／MI血漿。免疫保護(hù)馬在攻毒后3個(gè)T}J血漿病毒RNA降到很低水平，用實(shí)時(shí)定越PCR方法檢測不到。但在接個(gè)試驗(yàn)過程中，在INVIVOCHARACTERIZATIONSOFINFECTIOUSMOLECULARCLONEANDITSDERIVATIVESOFDONKEYLEUCOCYTEARENUATEDEQUINEINFECTIOUSANEMIAVIRUSABSTRACTINORDERTOINVESTIGATETHEMOLECULARMECHANISMOFTHEATTENUATIONANDPROTECTIONOFDONKEVLEUKOCYTEATTENUATEDEQUINEINFECTIOUSANEMIAVIRUSDLAEIAV，THECOMPLETEGENOMESOFDLA．EIAVANDEIAVSWAINLIANNINGLEIAVPROVIRUSESWERECLONEDANDSEQUENCED．THEINFECTIOUSMOLECULARCLONEPOK8266OFDLAEIAVWASCONSTRUCTANDITSDERIVEDVIRUSWASOBTAINEDDESIGNATEDASVOK8266，BASEDONPOKE266，ACHIMERICINFECTIOUSMOLECULARCLONEPOKVITRWASCONSTRUCTEDBYREPLACEDTHELONGTERMINALREPEATERRSEQUENCEOFDLAEIAVWITHTHECOUNTERPARTOFLEIAV,ANDITSDERIVEDVIRUSWASGENERATEDDESIGNATEDASVOKVLTR．INTHISSTUDY,SEVENEIAVNEGATIVEHORSESWERERANDOMLYDIVIDEDINTOFOURGROUPS．RWOHORSESF4AND5INGROUP1WEREINOCULATEDWITHVOKVLTR,TWOHORSES6AND7JNGROUP2WITHVOK8266，ONEHORSEINGROUP3WITHDLAEIAV,AND2HORSES1拌AND2INGROUP4WERESERVEDUNVACCINATEDCONTR01．EIGHTMONTHSPOSTINOCULATION，ALLHORSESWERECHALLENGEDWITHLEIAV．PREANDPOSTCHALLENGE，THEBODYTEMPERATUREOFALLHORSESWASRECORDED．ALLBORSESSHOWEDNOABNORMALCHANGEPREEHALLENGE,INDICATINGTHATALLVIRUSESUSEDWERESAFEFORHORSES，ANDTHEREPLACEMENTOFTHELTROFTHEATTENUATEDMOLECULARCLONEBYLEIAVLTRDIDN’TRESULTINVIRULENCEINCREASING．THEHORSESINGROUP4EXPERIENCED12TO41EPISODESOFFEVERANDDIED19DAYSAND134DAYSPOSTCHALLENGE，RESPECTIVELY．THEHORSESINGROUPS1AND3HADNOABNORMALCHANGESPOSTCHALLENGE，INDICATINGTHATTHECHIMERICVIRUSANDVACCINEVIRUSPROTECTEDTHEHORSESFROMCHALLENGE．INGROUP2．THEHORSE酬SHOWEDNOCLINICALSIGNS．BUTTHEHORSE7EXPERIENCED7EPISODESOFFEVERANDDIED185DAYSPOSTCHALLENGEDEADHORSES1撐，2撐AND7撐SHOWEDTYPICALEIAPATHOLOGICALCHANGESATAUTOPSY,ANDSURVIVEDHORSES4拌，5群，6群AND8WEREEUTHANIZED450DAYSPOSTCHALLENGE，ANDDISPLAYEDNOPATHOLOGICALCHANGES．THEANTIBODIESAGAINSTTHEE1AVS仇LCTURALPROTEINSP15，PL1，P26，P9ANDGP45WEREMEASUREDBYELISABASEDONTHERECOMBINANTPROTEINSEXPRESSEDINE．COLI．THEANTIBODYIEVELSINDUCEDBYP15，PL1ANDP9PROTEINSWEREVERYLOWORUNDETECTABLEPRECHALLENGE，ANTIBODYAGAINSTP26WASINCREASEDSOONAFTERIMMUNIZATIONANDSLOWLYCOMEDOWNLATER,ANTIBODYTOGP45WASINCREASEDSLOWLYANDPERSISTENTATHIGHLEVEL．AFTERCHALLENGE．THETITERSOFANTIBODIESAGAINSTTHEFIYESTRUCTURALPROTEINSINHORSES1，2AND7INCREASEDSIGNIFICANTLYANDSUSTAINEDTILLDEATH，ANFIBODI囂TOPL5，P26ANDGP45WEREALSOINCREASEDINHORSES6AND8．ANTIBODIESLEVELAGAINSTTHEFIVESTRUCTURALPROTEINSDIDNOTSHOWSIGNIFICANTCHANGEINHORSES4AND5AFTERCHALLENGE．THELOWEROI“NOREMEMBRANCERESPONSEMIGHTREFLECTGOODPROTECTION．THEANTIGP45TITERSHADNOOBVIOUSDIFFERENCEBETWEENDISEASEDANDPROTECTEDHORSES，SOTHEANTIGP45ANTIBODYSEEMSTOHAVENORELATIONSHIPWITHTHEDISEASEPROGRESS．THEEIAVPROVIRUSDNAINPBMCSANDTHEEIAVRNAINPLASMAOFINOCULATEDHORSESWEREⅢ【

簡介：河北農(nóng)業(yè)大學(xué)碩士學(xué)位論文羊附紅細(xì)胞體生物學(xué)特性研究及其PCR和ELISA檢測方法的建立姓名楊鵬華申請學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師秦建華20040618BIOLOGICALCHARACTERISTICSOFEPERYTHROZOONOVISANDDETECFIONOFUSINGTHEPOLYMERASECHAINREACTIONANDELISASPECIALITYPREVENTIVEVETERINARYMEDICMEPOSTGRADUATEDYANGPENGHUARESEARCHDIRECTIONANIMALPARASITOLOGYTUTORPMFQINJIANHUAABSTRACTTHREEKINDSOFANTICOAGULANTWEREUSEDINTHEBLOODINFECTEDBYEPERYTHROZOONOVISTHERESULTINDICATEDTHATCITRATEAFFECTEDIT’SMULTIPLICATIONMOSTINVITROANDEPERYTHROZOONOVISINCREASEDBYADDGLUCOSEANDNAHC03INCITRATEBLOODSAMPLEMUSTBETREATEDWITHIN12HAFTERCOLLECTFXOMTHEBODYEPIDEMIOLOGYINVESTIGATEDTHROUGHGIEMSASTAINEDBLOODSMEARSRESULTINDICATEDTHATSEASONALINFECTIONISACHARACTEROFTHISDISEASEANDARTHROPODSISAKEYTOITINFECTIONRATEINCREASEDFROMCOLDWINTERTOWARMANDDAMPSPRING80％SHEEPCANBEINFECTEDBYEPERYTHROZOONOVISINHOTSUMMERGENERALYTWOPRIMERSWERESYNTHESIZEDBASEDONTHECONSERVATIVESEQUENCEOFEPERYTHROZOONOVIS16SRRNAA1000BPFRAGMENTWASAMPLIFIEDBYUSINGPOLYMERASECHAINREACTIONTHEAMPLIFIEDFRAGMENTSWITHTHEEXPECTEDSIZEWEREIDENTIFIEDBYECORIRESTRICTIONDIGESTIONTHECROSSINGREACTIONANDSPECIFICREACTIONINDICATEDTHATPCRISQUICK，SENSITIVEANDSPECIMITISAEFFECTIVEWAYTOBEUSEDTODIAGNOSEEPERYTHROZOONOVISFROMTHEWHOLELEVELTHEOPTIMALCONDITIONOFELISAWASESTABLISHEDINTHISEXPERIMENTTHEDILUTIONOFANTIGEN，POSITIVESERUNLANDHRPGOATANTIRABBITIGGISL64，L160ANDL1000RESPECTIVELYTHEOPTIMALREACTIONTIMEOFANTIBODYANDHRPGNATANTIRABBITIGGISLHTHEMINIMUMDETECTIONOFEPERYTHROZOONOVISIS5030G／M1THECROSSINGREACTIONINDICATEDTHATELISAISAQUICK，SENSITIVEWAYADAPTTOCOLONYCHECKKEYWORDSEPERYTHROZOONOVIS；EPIDEMIOLOGY；ELISA；POLYMERASECHAINREACTION

簡介：摘要摘要以全展后的兩優(yōu)培九劍葉為研究對象，同時(shí)期的父本9311和母本培矮64S為對照，對兩優(yōu)培九、父本9311和母本培矮64S在衰老進(jìn)程中不同時(shí)期劍葉的光能轉(zhuǎn)化功能特性，膜脂脂肪酸成分變化及葉片超微結(jié)構(gòu)變化進(jìn)行了研究，揭示了劍葉整個(gè)衰老過程中細(xì)胞水平上的一些變化的規(guī)律。結(jié)果顯示一、三材料凈光合速率呈前期上升中后期下降。呼吸速率的變化為前期較低，中期稍有上升，但后期差異較大。兩優(yōu)培九在光能轉(zhuǎn)化上有一定的超親優(yōu)勢。兩優(yōu)培九的PSI、PSII電子傳遞活性在整個(gè)劍葉生長期間都是最高的，其次是父本，母本最低。從遺傳角度看，兩優(yōu)培九在光能轉(zhuǎn)化方面，更傾向于父本，而受母本影響則相對較小。二、膜脂脂肪酸不飽和指數(shù)是一個(gè)比較有意義的指標(biāo)。兩優(yōu)培九、父本9311和母本培矮64S不同部位脂肪酸不飽和指數(shù)呈起始和最后略低，中間稍高的趨勢。意味著在植物膜脂成分中，不飽和脂肪酸的含量對維持膜的正常生理功能有著重要的作用，飽和脂肪酸含量的提高往往不利于植物的生長發(fā)育，這可能與膜的流動(dòng)性能有關(guān)。整個(gè)劍葉生長期間，脂肪酸中140、160和18L的含量變化基本呈上升趨勢；而161、182和183則是呈現(xiàn)下降趨勢。膜脂脂肪酸不飽和指數(shù)的下降主要是由于上三種脂肪酸下降所致。兩優(yōu)培九三數(shù)據(jù)的變化比較平穩(wěn)。結(jié)構(gòu)上的穩(wěn)定意味著可以保持功能上的穩(wěn)定，兩優(yōu)培九可以在葉片水平上長時(shí)間的保持的凈光合速率及呼吸速率的穩(wěn)定與此有相當(dāng)大的關(guān)系。這為兩優(yōu)培九長時(shí)間的積累同化產(chǎn)物，提高最終產(chǎn)量提供了結(jié)構(gòu)上的保證。兩優(yōu)培九在膜臘脂肪酸成分上與雙親的遺傳關(guān)系與母本更為相似。三、細(xì)胞及葉綠體結(jié)構(gòu)觀測表明劍葉生長早期，細(xì)胞內(nèi)葉綠體多呈狹長形或橢圓形，數(shù)目較多。細(xì)胞核體積較小，核膜核孔清晰，核內(nèi)核質(zhì)濃密而均勻。隨著生長進(jìn)程的加深，葉綠體體積出現(xiàn)增大，內(nèi)部類囊體膜基粒片層部分模糊不清，淀粉粒變大，同時(shí)出現(xiàn)嗜鋨滴。但此時(shí)細(xì)胞核與線粒體沒有出現(xiàn)較大的變化。在劍葉衰老后期葉綠體已經(jīng)膨大，基粒片層松散紊亂，嗜鋨滴數(shù)目增多，體積增大，線粒體減少，內(nèi)部出現(xiàn)空泡化；細(xì)胞核的體積增大，核膜出現(xiàn)消融，核孔擴(kuò)大，核質(zhì)疏松，并向外散逸。兩優(yōu)培九在光合能力上有～定的結(jié)構(gòu)優(yōu)勢一、細(xì)胞內(nèi)葉綠體數(shù)目較多，基粒

簡介：中國農(nóng)業(yè)科學(xué)院博士學(xué)位論文小麥冰草異源二體附加系的細(xì)胞學(xué)和分子生物學(xué)檢測姓名王睿輝申請學(xué)位級(jí)別博士專業(yè)作物遺傳育種指導(dǎo)教師李立會(huì)20041015ABSTRACTMORPHOLOGICAL，CYT010西CAL，MOLECULARMETHODSANDPATHOIOGICALSCREENINGWEREEMPLOYEDTOANALYZET11ECYTOLOGICALSTABIL吼A磬0NOMICCHARACTEFSANDHOMOEOIOGOUSRELATIONSHIPBETWEENBREADWHEATANDWHEAT掣ASS口GR。刃，，0“CR括缸啪2N4X28CHROMOSOMEOF20WHEAT卅C，缸缸。堋CHROMOSOMEDISOMICADDITIONLLNESAM伽GTHE20ADDINONLINESA11ALYZED，GREATV耐ATIOLLSOFCYT0109ICALSTABILITIESEXISTEDPLA工1TSWITHCHROMOSOMECONSTIMTIONOF2N244V撕EDGREAⅡYBETWEENOPENPOLLINATEDPROGENIESOF10WHEAT“盯細(xì)臼聊LADDI石ONLIILES，RANGMG丘OM333％TO100％，W汕AVERAGE7478％INTHEREMAINING10ADDIDON1INES，NOPLANTSWITLL2N“WEREOBSERVEDWⅫEMEPLANTSWITH2N42PREDOMING把DHOWEVE‘GISHANALYSISFORPFOGENIESWIM2N42OF4DISOMICADDI廿0NLINESSHOWEDT11ATONETRANSLOCATIONLINEANDONESUBSTITLLTION1INEBETWEEN爿州J缸咖AILDWHEATCHROMOSOMESWERERECOVEREDNISSUGGESTEDTHATTRANSLOCATIONORSUBSTITILTIONBETWEEN4CR括缸咖ANDWHEATCLLROMOSOMESOCCU“℃DREAMLYTHEREFORE，S姍GTLLENINGDETECTIONFORTHEALIENCHROM撕NINTHEPROGENIESOFDISOMICADDITIONLINESC柚BEHELP“FORT11ERECOVERYOFTRANSLOCATIONORSUBSDTUTIONLI工1ESWITTLDESM出1EGENES‰爿C脅柳啪GENOMEACCORDINGTOMES曲ILARMESOFMORPHOLOGICAL鋤DAGRONOMICCH礎(chǔ)N℃FS，TENDISOMICADDITION1INESCALLBEDHIDED商。如EGROUPS，WIMWHICHEACHGROUPHASI協(xié)T，PICALCHARACTERISTICSFOFTHEPROGENIES塒THCHROMOSOMAJCOILSTINLTIONOF2N42，THEYDISPLAYEDTHICKERSTEMS，10NGERSPIKES，TALLERHEI曲TANDLONGERINTEMODELENGMUNDEMEATHSPIKETLLANMOSEOFTHECOUNTERPAR七PLA士1乜WITLL2N44OBVIOUSDINBRENCEOFSEEDCOLORWASOBSERVEDONTLLETWOTYPESOFPLALLTSWIMDIFFBRCNTCHROMOSOMALCONS吐TUTIONSSSRRESULTSSHOWEDT11ATSSRPRIMERSE乜丘OMWHEAT硎打C姍D即FFV“MAND爿曙F如卵細(xì)甜CAFF，CANAMPLIFIED一口由把N刪CH】0MOSOMESPECIFICBANDSFBM爿∥如缸抑ZGENOMEAMONG101POLYMORPHICPRIMEFSBETⅥ，EENFUKUHOANDZ559，WHICHARETHEWHEATAND』CR打缸F刪PARENTSRESPECTIVEU24PRJMERSETSCAN鋤PLIFYⅥMEA略RASSSPECIFICBANDS打OMTHEWHEAT爿柏缸F硼DISOMICADDITIONLINESWNHONESSRMARKERFROMEACHHOMOELOGOUSGROUP，THEHOMOEOLOGOUSRELATIONSHIPBETWEENWHEAT肌D彳酬SF謝姍2CHI講NOSOMESCANBEESTABLISHEDREADILYHOWEVELWITHTHEINCREASINGOFSSRMARKERS，SUCHHOMOEOLOGOUSREIATIONSH砸DISPLAYEDCOMPLIC蘸C曼零F彝＆￡愛妻霪C零霉L；I氍曼I嚏尊諱爿J蠹醣；N翼婪剿門掣匪尊ETI弱崩玎J。IG囂羹II；I堅(jiān)T藩“I簿L莩吐HR蒼Q叢N醐鼉G藩驀二TT孽墓鬟“F刪LM目KL』I鰣薹甜L；I耋劍I濘莖墜葉GIN∥虻Q垂墜蟄㈣竺麓鬃女』1上囂馘譬LUT如II；羹蘭S舔GI矗；H蜷L輯“藿￡∈蕞LI崮㈨LI警L贅U囊堅(jiān)“裂羈EM蓮衙簿L杵』U結(jié)∞IN蔓辮；；≠基鰉D9IO；囂FW；萋匱ERIMINATION0FBARLEYCLLROMOSOMESTHEOLAPPLGENET81285292174KOEBNERANDSHEPERD，1986175KNLSE，A1967IMERGENERICHYBRIDSBE鉚EEN脅礎(chǔ)堋W咖MLTSSP如出向啪VPALL粥2N_14AND＆C砒CER陽KLVPETKUS2N_14INRQ瑚，VE把R加∞7硎D昭疵礎(chǔ)啪F∞脅GP”甜6D曲

簡介：廣西大學(xué)碩士學(xué)位論文水牛卵巢卵泡顆粒細(xì)胞凋亡的生物學(xué)特征姓名李恭賀申請學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師何寶祥20050501BIOLOGICALCHARACTERISTICSOFBUFFALOOVARIANFOLLICULARGRANULOSACELLSAPOPTOSISABSTRAETGENERALLY，THEMOSTOFFOLLICLESUNDERGOATRESIAANDDEGENERATIONDURINGTHEIRDEVELOPMENTINMAMMALS，ANDONLYAFEWOFTHEMGROWUNIMERRUPTEDLYTOMATURATIONUNDLOVULATIONTHEMECHANISMOFTHEFOLLICLEATRISIASTILLREMAINSUNCLEARALTHOUGHMANYSTUDIESONTHEFOLLICLEATRESIAHAVEBEENCONDUCTEDBYMANYRESEARCHERSINTHISSTUDYSOMEEXPERIMENTSONFOLLICULARATRESIAWERECARRIEDOUTINBUFFALOASFOLLOWS1、THEMORPHOLOGICALCHANGESOFAPOPTOSISONTISSUESECTIONSOFHEALTHYFOLLICLESANDATRETICFOLLICLESINBUFFALO’SOVARIESWEREOBSERVEDBYLIGHTMICROSCOPEANDTRANSMITELECTRONICMICROSCOPE，THEAPOPTOSISOFTHEGRALULOSACELLSDREWFROMBUFFALOOVARIANFOLLICLEWASEXAMINEDBYFLUOROMICROSCOPETHERESULTSHOWNTHATFOLLICULARCELLSHAVETYPICALCHANGESOFAPOPTOSISSUCHAS，CONDENSATIONOFCHROMATIN，NUCLEOPLASMICSEGMENTATIONANDCONGREGATEDUNDERNUCLEARMEMBRANE；CELLMEMBRANESHRINKAGE，ORGANDIECONGREGATEANDWEREDARKSTAINEDUNDERMICROSCOPETHEREWEREAMASSOFAPOPTOTICBODIESWHICHWEREENWRAPPEDBYINTEGRATEMEMBRANEINATRESICFOLLICULARCAVITY；APOPTOTICGRANULOSACELLSDISPLAYEDBRIGHTERBLUETHANNORMALCELLSUNDERFLUOROMIEROSCOPE2、THE“SMEAR”PATTERNTHATREFLECTEDDNABREAKDOWNWASNOTFOUNDINHEALTHYFOLLICLESANDTHE“SMEAR”WASFOUNDINATRETICFOLLICLESBYGRANULOSACELLSDNAELECTROPHORETICANALYSISBUTTHETYPICAL“LADDER’’PATTEMOFGRANULOSAAPOPTOSISWASNOTFOUNDINBOTHHEALTHYFOLLICLESANDATRETICFOLLICLES3、AOPTOTICCHANGESOFGRANULOSACELLSWEREEXAMINEDINTHEHEALTHYANDATRETICTOLLIELESOFBUFFALO’SOVARIESBYTUNEL、THERESULTSSHOWEDTHATTHEREAREFEWTUNELPOSITIVECELLSINHEALTHYFOLLICLESBUTTHEREAREMANYTUNELPOSITIVECELLSINATRETICFOLLICLES，NAMELYLOTSOFDNAFRAGMENTSFROMAPOPTOFICCELLSORNECROTICONESCANBEDYEDINBLACKBROWN，BEINGPOSITIVE

簡介：四川農(nóng)業(yè)大學(xué)碩L學(xué)位論文小麥特殊材料R149的細(xì)胞生物學(xué)和分子生物學(xué)研究王翔指導(dǎo)教師廷止瞳越擐學(xué)科專業(yè)J目墅剛勤野生_一研兜方向鹽王細(xì)胞生塑生2005年5月ABSTRACTSTUDYONTHESPECIALWHEATMATIERIALR149BASEDONCYTOBIOLOGYANDMOLECULARBIOLOGYWANGXIANGPLANTGENETICSANDBREEDINGDIRECTEDBYPROF．RENZHENGLONGADVANCEDWHEATCULTIVARR149WHOSEPARENTSARECOPMONWHEAT伢ITICUMAESTLVUMANDRYESECALECEREALSISBREDBYNEWCHROMOSOMEENGINEERINGWAYS“THEUSEOFMONOSOMICRYEADDITION1INESFORTRANSFERRINGRYECHROMATININTOWHEAT”．USINGTHECBANDINGTECHNIQUE，GENOMEINSITUHYBRIDIZATION，ANDPROTEINSEPERATIONTECHNIQUEANDPCRSTUDIEDR149CULTIVARSYSTEMATICALLY．INORDERTOVERIFYTHETHEORY“INDUCTIONOFSMALL一SEGMENTTRANSLOCATION”．THESTUDIESSUBSTANTI8TEDFEASIBILITYOFTHETHEORY．1．THESTUDYRESULTSSUGGESTEDTHATTHEMONOSOMICADDEDRYECHROMOSOMESINWHEATCOULDINDUCETRANSLOCATIONBETWEENWHEATANDRYECHROMOSOMESEFFECTIVELYANDTHATTHEADDEDMONOSOMICCOULDINDUCESMAL1SEGMENTTRANSLOCATIONSSTRANSLOCATIONWITHHIGHFREQUENCY．THETHEORY“INDUCTIONOFSSTRANSLOCATION”ISSIGNIFICANTINPLANTBREEDINGCOMPAREWITHOTHERWAYSOFINDUCTIONOFTRANSLOCATION．2．THERESULTSOFTRADITIONALCYTOLOGYTECHNIQUESSHOWEDTHATR149HASANORMALCYTOLOGYBEHAVIORANDSTABLEGENETICSYSTEM．WHEREAS，THEPLANTSEXHIBITEDPHENOTYPICALTERATIONCOMPAREDWITHITSWHEATPARENT．3．THERESULTSOFTHEGISHINDICATEDTHATTHECONTROLMATERIALSSHOWEDALIENCHROMOSOMECLEARLYANDR149HADARELATIVEREGULARHYBRIDIZATIONSITEALONGTHESPECIALCHROMOSOMES．THEREFORE，WECOULDCONCLUDETHATITISPOSSIBLETHATR149HADSOMESEGMENTSOFTHERYECHROMOSOME．4．USINGTHEPRIMERSOFSPECIFICREPEATEDSEQUENCEFROMS．CEREALEAMPLIFIEDTHER149，THERESULTSSHOWEDTHATPSCLT9．1、PSCH20COULDAMPLIFIEDTHEPRODUCTSCORRECTLY．5．BASEDONSDSPAGEANALYSIS，R149SHOWEDNORYESPECIALGLUTENINPROTEIN．Ⅱ

簡介：山東農(nóng)業(yè)大學(xué)碩士學(xué)位論文抗AIV（HN）抗體獨(dú)特型雜交瘤細(xì)胞株的建立及其產(chǎn)物生物學(xué)特性的研究姓名彭軍申請學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師牛鐘相20050520英文縮寫AIAⅣMCABTCIDSOPDSOODHLAIMPAIHAHICFAIFARPMIGGHRP0PDPEGMW4000RP～Ⅱ16∞HMINSM英文全稱中英文縮略表AVIANINFLUENZAAVIANINFLUENZAVIRUSMONOCLONALANTIBODYMEDIANTISSUECULTUREINFECTIVEDOSEMEDIANPROTECTIVEDOSEOPTICALDENSITYHIGHLYPATHOGENNIEAVIANINFLUENZAMILDLYPATHOGENNICAVIANINFLUENZAHEMAGGLUTININHEMAGGLUTINININHIBITIONTESTCOMPLETEFREUNDSADJUVANTINCOMPLETEFREUND’SADJUVANTROTATIONPERMINUTEIMMUNOGLOBULINGHORSERADISHPEROXIDASEORTHOPHENYLENEDIAMINEPOLYETHYLENEGLYCOL4000HOURMIRTHTESECONDM臥ROLLTER中文名稱禽流感禽流感病毒單克隆抗體半數(shù)組織培養(yǎng)感染量半數(shù)保護(hù)量光密度高致病性禽流感低致病性禽流感血凝試驗(yàn)血凝抑制試驗(yàn)弗氏完全佐劑弗氏不完全佐劑每分鐘轉(zhuǎn)數(shù)免疫球蛋白G臘根過氧化物酶鄰苯二胺聚乙二醇RPMI1640培養(yǎng)基小時(shí)分鐘秒微升

簡介：中國農(nóng)業(yè)大學(xué)博士學(xué)位論文環(huán)境低溫誘發(fā)肉雞肺血管重塑的細(xì)胞生物學(xué)機(jī)制研究姓名王建琳申請學(xué)位級(jí)別博士專業(yè)基礎(chǔ)獸醫(yī)學(xué)指導(dǎo)教師喬健20060501重要因素，這兩方面是環(huán)境低溫誘發(fā)肉雞肺血管重塑的主要細(xì)胞生物學(xué)機(jī)制。II關(guān)鍵詞肉雞肺動(dòng)脈高壓綜合征環(huán)境低溫細(xì)胞增殖原癌基因肉雞

簡介：廣西大學(xué)碩士學(xué)位論文奶牛附紅細(xì)胞體病PCR檢測方法的建立及其病原生物學(xué)初步研究姓名陳明申請學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師黃維義張為宇20060701／奶牛附紅細(xì)胞體病PCR檢測方法的建立及其病原生物學(xué)初步研究特異性引物，建立奶牛附紅細(xì)胞體病的POR診斷方法。特異性試驗(yàn)和敏感性試驗(yàn)表明，該診斷方法與豬肺炎支原體、雞毒支原體、大腸桿菌、腸道沙門氏菌、葡萄球菌、雞艾美耳球蟲、牛雙芽巴貝斯焦蟲無交叉反應(yīng)；能檢測的奶牛附紅細(xì)胞體陽性血樣DNA最低濃度為0154NG／P_L，同時(shí)能檢測出在40存放長達(dá)三個(gè)月的血樣。通過臨床血樣檢測，證明該方法可以用于本病的診斷。另外建立了奶牛附紅細(xì)胞體病和伊氏錐蟲病的二重POR診斷方法，其擴(kuò)增片段大小分別為415BP和227BP。特異性試驗(yàn)和敏感性試驗(yàn)表明，該方法與豬肺炎支原體、大腸桿菌、葡萄球菌、雞艾美耳球蟲、牛雙芽巴貝斯焦蟲無交叉反應(yīng)；能檢測的奶牛附紅細(xì)胞體陽性血液DNA最低濃度和伊氏錐RLLFL’ART液DNA最低濃度分別為043NG／“L和0106NG／LAL。臨床試驗(yàn)表明，該方法可用于奶牛附紅細(xì)胞體病和伊氏錐蟲病的診斷。為了初步探索不同種動(dòng)物附紅細(xì)胞體之間是否存在交叉感染，本研究首先摸索體外培養(yǎng)水牛紅細(xì)胞的最佳條件，然后用染色鏡檢為附紅細(xì)胞體陽性的人、奶牛、小白鼠、狗、貓和兔紅細(xì)胞在體外感染陰性的水牛紅細(xì)胞。試驗(yàn)結(jié)果表明當(dāng)PH值為72，C0為6％，1640為基礎(chǔ)培養(yǎng)，含胎牛血清40％的完全培養(yǎng)基可用于水牛紅細(xì)胞的培養(yǎng)，在體外可以成功培養(yǎng)長達(dá)30天；在體外交叉感染試驗(yàn)中，奶牛和貓的附紅細(xì)胞體最易感染水牛，其感染率可達(dá)60％以上，其次是小鼠和狗，其感染率在3040％之間，最低為兔，感染率只能達(dá)到15％，而人的附紅細(xì)胞體在體外不能感染水牛紅細(xì)胞。本研究通過對不同種動(dòng)物附紅細(xì)胞體血液涂片染色、體外培養(yǎng)及體外

簡介：分類號(hào)密級(jí)石河子大學(xué)碩士學(xué)位論文綿羊附紅細(xì)胞體的生物學(xué)檢測及其對紅細(xì)胞、T淋巴細(xì)胞影響的研究研究生王麗指導(dǎo)教師申請學(xué)位門類級(jí)別學(xué)科、專業(yè)名稱研究方向所在學(xué)院動(dòng)物傳染病的診斷與防治動(dòng)物科技學(xué)院中國新疆石河子2006年3月但藥物聯(lián)合治療綿羊附紅細(xì)胞體病具有更明顯的效果。附紅細(xì)胞體感染羊紅細(xì)胞的生理指標(biāo)、免疫指標(biāo)結(jié)果顯示隨感染程度的增加，以紅細(xì)胞總數(shù)減少、血紅蛋白濃度HBG和紅細(xì)胞壓積PCV、SOD酶活力和ATPASE酶活性顯著降低為主要指征，出現(xiàn)嚴(yán)重的貧血癥。紅細(xì)胞酵母花環(huán)EC3BRR花環(huán)數(shù)在實(shí)驗(yàn)中隨羊附紅細(xì)胞體感染程度的增加而下降、而紅細(xì)胞免疫復(fù)合物花環(huán)EICR花環(huán)數(shù)在隱性組表現(xiàn)下降，急性感染組中表現(xiàn)回升的趨勢，說明紅細(xì)胞在抗附紅細(xì)胞體感染中起了一定作用，但由于紅細(xì)胞數(shù)量的減少，使參與機(jī)體非特異反應(yīng)的紅細(xì)胞免疫功能降低?；钚訲淋巴細(xì)胞在酸性非特異性醋酸萘酯酶檢測中呈下降趨勢。綜合分析表明羊附紅細(xì)胞體感染不會(huì)引起較強(qiáng)的細(xì)胞免疫，但會(huì)使紅細(xì)胞免疫功能降低，從而減弱了機(jī)體抵抗外界感染的能力。本研究為病原學(xué)鑒定、本病的預(yù)防和控制提供了理論依據(jù)。關(guān)鍵詞綿羊附紅細(xì)胞體透射電鏡PCR；紅細(xì)胞T淋巴細(xì)胞；免疫功能論文類型A理論研究B應(yīng)用研究Ⅱ

簡介：內(nèi)蒙古農(nóng)業(yè)大學(xué)碩士學(xué)位論文大骨雞、青殼蛋雞成纖維細(xì)胞庫的建立及生物學(xué)特性研究姓名躍華申請學(xué)位級(jí)別碩士專業(yè)動(dòng)物遺傳育種與繁殖指導(dǎo)教師道爾吉關(guān)偉軍20060501RESEARCHONTHEESTABLISHMENTANDCHARACTERIZINGOFDAGUCHICKENANDQINGKECHICKENFIBROBLASTCELLLINESABSTRACTEMBRYOSOFDAGUCHICKENANDQINGKECHICKENWERECULTIVATEDBYDIRECTADHERENTCULTUREMETHODSTOCULTUREFIBROBLASTANDTHENTHEEELLLINESWEREFROZENINLIQUIDNITROGENTOCONSERVATIONTHECELLSWEREPASSAGEDNOMORETHAN4GENERATIONS，ANDTHECELLFREEZINGDENSITYISABOUT2～3X100CELL／ML，DAGUCHICKENANDQINGKECHICKEN’SSAMPLENUMBERSOFEACHKINDARE19AND9，CONSERVEDNUMBERSARE91AND42AMPULES，BIOLOGICALCHARACTERSOFFIBROBLASTWEREANALYSEDACCORDINGTOTHEREQUESTOFCONSTRUCTINGCELLLINETHERESULTSWEREASFOLLOWS1CULTUREDCELLS’MORPHAISTYPICALFIBROBLAST，WHICHISFUSIFORMATEORUNREGULARTRIANGLE，CELLSISRADIANT，F(xiàn)LAMINGORROTATIVEDURINGGROWINGTHROUGHMICROSCOPETHEMORPHADIVERSITYOFFIBROBLASTINDIFFERENTBREEDCANBEOBSERVED2THEGROWTHCURVEOFCULTURECELLINVITROAPPEARED“S”SHAPEANDILLUSTRATEDTHATCELLSGROWINGINCLUDEDLATENTPHASE，LOGARITHMICGROWTHPHASEANDSTAGNATEPHASE3THERESULTOFCELLSURVIVALRATEDESCENDINGSHOWEDTHATCELLSWEREINJUREDDURINGCRYOPRESERVATIONBUTIT’SSURVIVALRATIOSURPASSED90％ADHERENTCELLAFTERREVIVALGROWEDASFASTLYASCELLBEFOREFREEZINGTYPEOFCELLISABSOLUTEFIBROBLAST4THERESULTOFMICROBIALDETECTIONSHOWEDTHATBACTERIA，F(xiàn)UNGI，VIRUSANDMYCOPLASMADIDNOTCONTAMINATECULTUREDCELLS5THERESULTOFCHROMOSOMEANALYSISTHECHROMOSOMEDIPLOIDNUMBEROFDAGUCHICKENANDQINGKECHICKENIS2N78，THERESULTSHOWEDCELLSSTILLKEPTDIPLOIDCONDITIONANDTHEHEREDITARYCHARACTERISSTABLETHEREISNOCROSSEDCONTAMINATIONBETWEENCELLLINES6THERESULTSINDICATETHATLIPOFECTINCOULDEFFECTIVELYTRANSFERPECFPN1PLASMIDINTOFIBROBLAST，F(xiàn)ORWHICHTHEOPTIMIZATONTRANSFECTIONCONDITIONSINVOLVEINFIBROBLASTGROWTHSTATE，DNA／LIPOFECTINMIXINGCONCENTRATIONRATIO，ANDEXPOSURETIMEOFFIBROBLASTTODNALIPOFECTINCOMPLEXESTRANSFECTIONEFFICIENCYOFDAGUCHICKENANDQINGKECHICKENWERE28％AND32％7THROUGHANALYZINGTWOKINDSOFISOENZYME，MDHANDLDH，DIFFERENTBREEDSHASSPECIFICBRANDSTHEREISNOCROSSEDCONTAMINATIONBETWEENCELLLINESTHECELLLINESTHATCONSTRUCTEDINTHISSTUDYACCORDWITHTHEREQUESTOFATCCWESUCCESSFULLYCONSTRUCTEDSEVENKINDSOFLIVESTOCKANDPOULTRYBREEDS’FIBROBLASTLINESANDCONSERVEGENETICRESOURCE