apob100介导的tpa基因在鸡体内表达及其在卵黄中沉积的研究word格式论文.docxVIP

摘要表达的融合蛋白均有生物学活性；通过对心脏、肝脏、血管、输卵管、肌肉、肾脏、等不同的组织免疫组化检测，发现融合蛋白在肝脏、肌细胞分布较多。apo-B100介导的t-PA融合基因在鸡体内的成功表达，表明1479bp的apo- B100介导的外源基因表达产物能够透过卵黄膜实现在卵黄中的沉积，证明了外源基因表达产物在卵黄中积淀这一途径的可行性。这一途径有望使鸡蛋成为预防心血管疾病的功能性食品，使鸡体成为药物蛋白开发生产的重要资源。关键词：组织型纤溶酶原激活剂；极低密度值蛋白；肝细胞；转基因；卵黄论文类型：应用基础研究II摘要Subject:ExpressionandactivitydetectionofApo-tPAfusionproteininchickenanditsaccumμLationinyolkSpecialty: BasicVeterinaryScienceName:YungongFengSupervisor:YinjuLiASSOCIATEPROFESSORPABSTRACTHumantissue-typeplasminogenactivator(t-PA),aspecific,fibrin-selective thrombolyticagentwasoneofthemosteffectivedrugstopreventandtreatthe thromboticdiseases.Apolipoproteinapo-B100wasnecessaryfortheformationand assemblesofverylowdensitylipoprotein(VLDLy).Duringthecourseofdeposition oforganicmatterintheyolk,VLDLywasswallowedintogrowthyolkoocytesinthe formofcompletelipoproteinparticles.Todeterminethefeasibilityabouttheaccum- ulationoft-PAintheeggyolkanddevelopthefunctionaleggsforanticardiovascular disease,theexpressionvectorwasconstructedbymeansoffusethefu-nctional?t-PA fragmentintneandapo-B100.Afteridentificationthecorrectexpressionfram- eworkinthemodelofhepatocyteinvitro,theexpressionvectorwastransfectedinto thechickenbyliposome.Whichthestudyoftheexpressionofthefusionproteininthe chickenanddepositionintheeggyolkpaveanroadontheuseofpoultryasabior- eactor to produce t-PA.ThetotalRNAoflayerliverwasextractedbymolecularbiologytechniquesin thisstudy.Theapo-B100geneclonedusingspecificprimersbyRT-PCRwasfused withthe?t-PA,functionalarea fragmentt-PAvialinkerprotein.Itwasclonedinto downstreamofCMVpromoterofpCDNA3expressionvectorandobtainedpCDNA3- apo-?tPAexpressionvector.Thentheexpressionvectorwastransfectedintothehep- atocytetoanalyzethecorrectreadingframe.Theexpressionleveloffusingprotein wasdetectedbyELISA.Thebiologicalactivityoftheexpressionproductwasdetected bychromognicsubstratemethods.Theexpressionvectorpackagedwithliposomewas injectedinto chickenbyintravenousinjection.Theeggwascollectedatdifferenttime. Theexpressionproductwasdet