本文選題：菌體蛋白 + 多肽��； 參考：《南京農(nóng)業(yè)大學(xué)》2006年碩士論文

【摘要】：接種卡介苗是當(dāng)今各國(guó)預(yù)防結(jié)核病最有效的手段之一。分支桿菌屬的成員如草分支桿菌、母牛分支桿菌等也被廣泛的應(yīng)用于疫苗和佐劑的制備。但是，由于是全菌制劑，，菌體中對(duì)免疫功能無作用、甚至起抑制作用的成分同時(shí)存在在疫苗中，卡介苗使結(jié)核菌素的皮試出現(xiàn)陽性反應(yīng)，因而限制了它們的應(yīng)用。 本文從結(jié)核分支桿菌中提取菌體蛋白和多肽，通過生物化學(xué)的方法對(duì)其進(jìn)行定量檢測(cè)。經(jīng)過動(dòng)物試驗(yàn)，探討分支桿菌菌體蛋白和多肽對(duì)雞特異性免疫和非特異性免疫的效應(yīng)，最終篩選出一種低毒、高效的新型免疫增強(qiáng)劑并確定了臨床最佳用藥劑量。 利用超聲波低溫破碎技術(shù)，提取、制備結(jié)核分支桿菌中的菌體蛋白和多肽，應(yīng)用生物化學(xué)方法(lowry法)測(cè)定菌體蛋白和多肽的含量；Tricine-SDS-PAGE電泳分析菌體蛋白組分。試驗(yàn)結(jié)果表明：不同批次提取的菌體蛋白含量相對(duì)穩(wěn)定，沒有明顯差異；組分分析結(jié)果表明，菌體蛋白由多種組分構(gòu)成，分子量由66kD到6.5kD不等。 按人常用劑量，根據(jù)人／雞體表面積比，計(jì)算雞用藥量。利用所提取的結(jié)核分支桿菌菌體蛋白和多肽投服試驗(yàn)雞。210只雞被均分為分支桿菌菌體蛋白高劑量組(0.9mg／只)、分支桿菌菌體蛋白中劑量組(0.45mg／只)、分支桿菌菌體蛋白低劑量組(0.225mg／只)、透析多肽組(0.3mg／只)、未破碎全菌體組(0.3g／只)、分支桿菌菌體蛋白注射組(0.45mg／只)和對(duì)照組(生理鹽水)七個(gè)組。除注射組外，其余六組均采用口服給藥。各組試驗(yàn)雞于給藥后7d、14d、21d、28d分四次撲殺，分別檢測(cè)雞外周血淋巴細(xì)胞轉(zhuǎn)化率、雞新城疫抗體滴度、紅細(xì)胞免疫水平、外周血淋巴細(xì)胞CD_3~+亞群和淋巴器官的組織發(fā)育情況。試驗(yàn)結(jié)果表明，分支桿菌菌體蛋白投服試驗(yàn)組均能提高雞體的特異性免疫和非特異性免疫功能，尤以菌體蛋白注射組和菌體蛋白中劑量口服組誘導(dǎo)機(jī)體免疫效果為好。
[Abstract]:Bacillus Calmette Guerin is one of the most effective means of preventing tuberculosis in various countries. Members of the genus Mycobacterium, such as Mycobacterium tumefaciens and Mycobacterium cow, are also widely used in the preparation of vaccines and adjuvant. However, because of the whole bacteria preparation, there is no effect on the immune function in the bacteria, and even the components of inhibition are present in the vaccine. BCG showed positive reactions to the skin test of tuberculin, thus limiting their application.
In this paper, bacterial protein and polypeptide were extracted from Mycobacterium tuberculosis and tested by biochemical method. After animal experiments, the effect of Mycobacterium protein and polypeptide on specific immunity and non specific immunity of chicken was investigated. Finally, a new type of low toxic and efficient immunosuppressive agent was selected and the clinical best was determined. A good dose of medicament.
The bacterial protein and polypeptide in Mycobacterium tuberculosis were extracted and prepared by ultrasonic low temperature crushing technology. The contents of bacterial protein and polypeptide were measured by the biochemical method (Lowry method). The bacterial protein components were analyzed by Tricine-SDS-PAGE electrophoresis. The results showed that the protein content of different batch of bacterial proteins was relatively stable and no obvious. The results of component analysis showed that the bacterial protein was composed of many components, with molecular weights ranging from 66kD to 6.5kD.
According to the common dose of human / chicken body surface area ratio, the amount of chicken medicine was calculated. The.210 chickens were divided into high dose group (0.9mg / only), the dose group of Mycobacterium body protein (0.45mg / only), and the low dose of Mycobacterium body protein in the Mycobacterium tumefaciens. Group (0.225mg / only), dialysis polypeptide group (0.3mg / only), unbroken whole bacteria group (0.3g / only), Mycobacterium tumefaciens protein injection group (0.45mg / only) and control group (physiological saline) seven groups. Except for the injection group, the other six groups were taken oral administration. Each group of test chickens was given 7d, 14d, 21d, 28d, four times after administration, respectively, to detect the peripheral blood drenching of chickens. The rate of Ba cell transformation, the titer of the antibody to the Newcastle disease, the level of the red cell immunity, the CD_3~+ subgroup of the peripheral blood lymphocyte and the tissue development of the lymphoid organs. The experimental results showed that the specific immune and non specific immune function of the chicken body could be improved by the experimental group of Mycobacterium tumefaciens, especially in the mycelial protein injection group and the mycelial egg. The white medium dose group had good immune effect.

【學(xué)位授予單位】：南京農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R392

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