本文選題：幽門螺桿菌　切入點(diǎn)：空泡毒素　出處：《重慶醫(yī)科大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

【摘要】： 第一部分幽門螺桿菌融合蛋白VacA-HpaA卵黃抗體的制備 目的:采用基因工程技術(shù)大量表達(dá)幽門螺桿菌(Helicobacter pylori, H.pylori)細(xì)胞空泡毒素(vacuolating cytotoxin antigen A,VacA)與粘附素(Helicobacter pylori adhesin A ,HpaA)的融合蛋白(VacA-HpaA),鑒定其抗原性與免疫原性后,以此融合蛋白作為抗原,制備高效價(jià)的VacA-HpaA IgY。 方法:通過(guò)大量培養(yǎng)重組菌pQE30-VacA-HpaA-DH5a,誘導(dǎo)表達(dá)獲得融合蛋白VacA-HpaA Ni2+-NTA樹脂純化。用Western blot鑒定融合蛋白VacA-HpaA對(duì)VacA和HpaA的抗原特異性,用融合蛋白免疫兔,獲得多克隆抗血清,采用間接ELISA法檢測(cè)其分別針對(duì)VacA和HpaA的免疫原性。將純化的融合蛋白免疫雞,水稀釋結(jié)合氯仿有機(jī)沉淀法提取IgY。ELISA法測(cè)定抗體產(chǎn)生的時(shí)間-效價(jià)變化。硫酸銨沉淀法純化濃縮VacA-HpaA IgY,SDS-PAGE分析純度,Bradford法測(cè)定蛋白含量,Western blot檢測(cè)其分別對(duì)VacA和HpaA的特異抗原性,ELISA法檢測(cè)效價(jià)。
[Abstract]:Preparation of VacA-HpaA yolk antibody of Helicobacter pylori fusion protein. Objective: to express the fusion protein VacA-HPA of Helicobacter pylori (H.pylori) vacuolating cytotoxin antigen AcaA) by genetic engineering, and identify its antigenicity and immunogenicity by using the fusion protein VacA-HpaA, which is a fusion protein of Helicobacter pylori adhesin A (HpaA), and to identify the antigenicity and immunogenicity of the fusion protein. Preparation of high titer VacA-HpaA IgY. Methods: the fusion protein VacA-HpaA Ni2-NTA was purified by mass culture of pQE30-VacA-HpaA-DH 5a. The antigenicity of the fusion protein VacA-HpaA to VacA and HpaA was identified by Western blot, and the polyclonal antiserum was obtained by immunizing rabbits with the fusion protein. The immunogenicity of VacA and HpaA was detected by indirect ELISA method. The purified fusion protein was used to immunize chickens. Water dilution combined with chloroform organic precipitation method was used to extract IgY.ELISA to determine the time-titer change of antibody production. Purification and concentration of VacA-HpaA IgYPS-PAGE by ammonium Sulfate precipitation Purity determination of protein content by Bradford method and Western blot Detection of its specific Antibody to VacA and HpaA. The titer was detected by Elisa.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R392

【引證文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 王R，

本文編號(hào)：1565532