本文選題：幽門(mén)螺旋桿菌 + CagA基因�。� 參考：《浙江大學(xué)》2006年碩士論文

【摘要】： 幽門(mén)螺桿菌是慢性胃炎、消化性潰瘍的病原體，也與胃腺癌及黏膜相關(guān)性淋巴瘤的發(fā)生密切相關(guān)。各國(guó)人群中Hp感染率約為50％，發(fā)展中國(guó)家超過(guò)70％。世界衛(wèi)生組織國(guó)際癌癥研究機(jī)構(gòu)已將該菌納入第一致癌原。CTB是一個(gè)有效的載體分子，抗原分子與CTB結(jié)合后，，與腸相關(guān)淋巴組織(Gut—associated lymphoid tissues，GALT)細(xì)胞表面的神經(jīng)節(jié)苷脂GM_1具有較強(qiáng)的親和力，這就更利于抗原的傳遞和免疫識(shí)別，引起口服耐受。番茄是基因工程中最為廣泛使用的模式植物之一并且番茄的果實(shí)可以直接實(shí)用是研制植物口服疫苗的理想植物體。 本研究構(gòu)建了含有融合基因CTB—UreB的果實(shí)特異啟動(dòng)子植物表達(dá)載體，并將已構(gòu)建好的含有融合基因CTB—CagA、CTB—UreB的植物表達(dá)載體，采用農(nóng)桿菌介導(dǎo)的葉盤(pán)轉(zhuǎn)化法，將融合基因轉(zhuǎn)入番茄中，為利用生物反應(yīng)器研制口服型植物疫苗奠定了先行基礎(chǔ)。 研究結(jié)果表明如下： ①利用農(nóng)桿菌介導(dǎo)的遺傳轉(zhuǎn)化系統(tǒng)將融合基因轉(zhuǎn)入番茄基因組中，經(jīng)過(guò)共培養(yǎng)和多次抗性篩選、誘導(dǎo)不定芽的分化和生根，得到了具有抗性的再生植株。再生植株的抗性芽分化率達(dá)40～60％。 ②PCR和RT—PCR驗(yàn)證目的基因已經(jīng)整合到了番茄的基因組中，PCR陽(yáng)性率＞70％。 ③篩選出來(lái)的陽(yáng)性植株繼續(xù)培養(yǎng)待果實(shí)成熟后取種子(F_1代)，將F_1代種子播種培養(yǎng)，經(jīng)過(guò)檢測(cè)所播種的第二代植株中陽(yáng)性率＞90％。 ④構(gòu)建了番茄果實(shí)特異啟動(dòng)子表達(dá)載體，經(jīng)酶切檢驗(yàn)正確后可用于轉(zhuǎn)化番茄外植體。
[Abstract]:Helicobacter pylori is the pathogen of chronic gastritis, peptic ulcer and gastric adenocarcinoma and mucosal associated lymphoma. The infection rate of HP is about 50% in the population of different countries and more than 70% in the developing countries. The World Health Organization's International Agency for Cancer Research has incorporated the bacterium into the first carcinogen. CTB is an effective vector molecule, and the antigen molecule binds to CTB. GM-1 has strong affinity with GM1 on the surface of Gut-associated lymphoid tissue (GALT) cells, which is more favorable for antigen transmission and immunological recognition, resulting in oral tolerance. Tomato is one of the most widely used model plants in genetic engineering. In this study, the fruit specific promoter plant expression vector containing CTB-UreB was constructed, and the plant expression vector containing CTB-CagAN-CTB-UreB was constructed by Agrobacterium tumefaciens-mediated leaf disk transformation. The transfer of the fusion gene into tomato lays the foundation for the development of oral plant vaccine by bioreactor. The results are as follows: 1 the fusion gene was transferred into tomato genome by Agrobacterium tumefaciens mediated genetic transformation system. The resistant regenerated plants were obtained by inducing the differentiation and rooting of adventitious buds. The resistant bud differentiation rate of regenerated plants reached 400.2The PCR and RT-PCR confirmed that the target gene had been integrated into tomato genome and the positive rate of PCR was > 70. (3) the selected positive plants continued to be cultured after fruit ripening, and the seeds were sowed and cultured in the first generation. The expression vector of tomato fruit specific promoter was constructed by detecting the positive rate of the second generation plantlets sown with a positive rate of > 90.4, which can be used to transform tomato explants by enzyme digestion test.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：Q943.2;R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 姜云水,方平楚,付亞萍,朱誠(chéng);幽門(mén)螺桿菌CagA蛋白與CTB融合基因的克隆及其植物表達(dá)載體的構(gòu)建[J];科技通報(bào);2004年03期

2 郭曉臨;王立娥;王蘭;董明;袁媛;;胃癌高發(fā)區(qū)高危人群血清HpCagA毒素相關(guān)蛋白檢測(cè)的意義[J];世界華人消化雜志;2001年05期

3 楊衛(wèi)紅,林三仁,金珠,王立新,丁士剛;幽門(mén)螺桿菌致病因子與胃粘膜病理變化的關(guān)系[J];中華消化雜志;2000年01期

4 杜奕奇,許國(guó)銘,紀(jì)徐淮,丁華,張洪富,滿(mǎn)曉華,孫振興;細(xì)胞毒素相關(guān)抗原(cagA)基因在中國(guó)人幽門(mén)螺桿菌中的分布及其臨床意義[J];中華消化雜志;1999年03期

5 嚴(yán)杰;幽門(mén)螺桿菌基因工程疫苗的研究[J];浙江大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2003年01期

6 關(guān)R

本文編號(hào)：2084622