哺乳运动朊蛋白叠朊的原核表达及牛朊蛋白单抗制备.pdf

博十后1．{1j报告 中文捕蜚 中文摘要 本文以基因克隆、表达和淋巴细胞杂交瘤技术为基础，制餐并签定了 针对秦川黄牛成熟朊蛋白mPrP的单抗，以期用于疯牛病的免疫诊断研究。 检测抗体的Western 毒PrP“的蛋白酶K抗性。 法纯化的腹水抗体。结果表明获得了7个能稳定分泌挽黄牛mPrP的细胞 示单抗N15，N25，N59，N63和N67i』；：j别相同的抗原表位，单抗N17和N64 识别mPrP羧基端的小矧表位。WesternBlot分析表明腹水单抗N64彝I 疯牛病的核心诊断试剂，为开发相关诊颐试剂盒奠定，基础。 关键词 疯牛病；脱蛋白：朊病毒：叠朊：单克降抗体；诊断试剂 博十后1．作报告 英文摘要 Abstract Monoclonal mature antibodies(mAbs)againstprion cattlewere andidentifiedonthebasisof Qinchuanyellow prepared gene and and mAbswere cloningexpression hybridomatechnology．Those toworkinthe ofmadCOWdisease． expected immunologicaldiagnosis TheORFsof and of prionprotein(PrP)genegeneQinchuancattle， Dpl nativebom ofGansuProvinceandtheHanmanwere PCR sheep acquiredby methods．Thenthe recombinant maurePrP expressionplasmidscontaining andmature wereconstructed．Thethreefusion (mPrP)geneDpl(mDpl)gene ofmPrPand were inthe Ecoli proteins mDpl overexpressedcorresponding induced IPTG．WesternBlot withmAbSAF-70asthe by analysis showedthatthethreefusionmPrPsexhibitieda antibody good andformeddimersinthe of K reactiongenic