本文關(guān)鍵詞：細(xì)粒棘球蚴原頭蚴雙向電泳圖譜數(shù)據(jù)庫的建立及標(biāo)記蛋白的定位　出處：《寧夏醫(yī)科大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 原頭蚴 蛋白質(zhì)組學(xué) 雙向電泳圖譜 蛋白靶標(biāo)

【摘要】：目的建立細(xì)粒棘球蚴原頭蚴雙向電泳的圖譜數(shù)據(jù)庫;用課題組前期工作得到的細(xì)粒棘球蚴原頭蚴三種重組抗原制備抗血清,純化出特異性抗體,以此識(shí)別雙向電泳圖中的蛋白質(zhì)位點(diǎn),為后期細(xì)粒棘球蚴原頭蚴蛋白質(zhì)組研究中蛋白質(zhì)的定位、蛋白位置的描述等原頭蚴雙向電泳圖譜的應(yīng)用奠定基礎(chǔ)。 方法1樣本制備:以細(xì)粒棘球絳蟲原頭蚴為樣品;2蛋白分離:應(yīng)用雙向電泳技術(shù)對(duì)其蛋白質(zhì)進(jìn)行分離;3圖像采集保存及分析:GS-800掃描儀采集圖像,通過bio-rad公司的PDquest軟件對(duì)得到的原頭蚴雙向電泳圖譜進(jìn)行分析,初步得出構(gòu)成原頭蚴的蛋白質(zhì)數(shù)量以及蛋白質(zhì)的分子量和等電點(diǎn)數(shù)據(jù);4表達(dá)純化:用細(xì)粒棘球蚴原頭蚴的三種重組蛋白免疫新西蘭家兔制備抗血清,將抗體純化后進(jìn)行western blot分析,識(shí)別原頭蚴雙向電泳圖譜中的已知蛋白質(zhì),利用這些已知蛋白作為圖譜中的標(biāo)記。 結(jié)果在原頭蚴雙向電泳圖中,分離出240個(gè)蛋白質(zhì)位點(diǎn),通過軟件分析初步得到了每個(gè)位點(diǎn)的分子量和等電點(diǎn)信息;成功的純化出三種細(xì)粒棘球蚴原頭蚴重組蛋白zw-5,14-3-3,p-29,并采用免疫學(xué)的方法制備出抗體,完成了抗體對(duì)原頭蚴蛋白質(zhì)組雙向電泳圖譜的識(shí)別。 結(jié)論1初步建立了細(xì)粒棘球蚴原頭蚴的雙向電泳圖譜;2在雙向電泳圖譜的基礎(chǔ)上,通過軟件分析得到了240個(gè)蛋白質(zhì)位點(diǎn),并分析獲得了這些蛋白質(zhì)的分子量和等電點(diǎn)信息;3利用原頭蚴重組蛋白制備抗體,在細(xì)粒棘球蚴原頭蚴的雙向電泳圖譜上標(biāo)記了三個(gè)已知蛋白靶標(biāo)。
[Abstract]:Objective to establish a two dimensional electrophoresis map database of Echinococcus granulosus (Echinococcus granulosus). The antiserum was prepared from three recombinant antigens of echinococcus granulosus, and the specific antibody was purified to identify the protein sites in the two-dimensional electrophoretogram. The results laid a foundation for the application of two-dimensional electrophoretic patterns of the proteome of echinococcus granulosus in the study of proteome, such as protein location and protein position description. Methods 1 sample preparation: Echinococcus granulosus was used as sample; (2) protein separation: the protein was separated by two-dimensional electrophoresis. 3Image acquisition, preservation and analysis: GS-800 scanner was used to collect images, and the two-dimensional electrophoresis patterns of protocercaria were analyzed by PDquest software of bio-rad Company. The data of protein quantity, molecular weight and isoelectric point of protein were obtained. 4 expression and purification: three kinds of recombinant proteins of Echinococcus granulosus were used to immunize New Zealand rabbits to prepare antiserum. The antibody was purified and analyzed by western blot. The known proteins in the two-dimensional electrophoretic map of the protocercaria were identified and used as markers in the map. Results 240 protein loci were isolated from the electrophoretogram, and the molecular weight and isoelectric point information of each locus were preliminarily obtained by software analysis. Three recombinant proteins zw-514-3-3p29 were successfully purified and antibody was prepared by immunological method. The antibody was used to recognize the two-dimensional electrophoretogram of protocercaria proteome. Conclusion 1Two-dimensional electrophoretic pattern of echinococcus granulosus was preliminarily established. 2 on the basis of two-dimensional electrophoresis map, 240 protein sites were obtained by software analysis, and the molecular weight and isoelectric point information of these proteins were obtained. 3. Three known protein targets were labeled on the electrophoretogram of E. granulosus by using recombinant protein of E. granulosus.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R383

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