本文關(guān)鍵詞： 霍亂毒素B亞單位 胰島素原 胰島素B鏈 大腸桿菌 轉(zhuǎn)基因馬鈴薯　出處：《寧夏大學(xué)》2005年碩士論文　論文類型：學(xué)位論文

【摘要】：口服胰島特異的自身抗原可誘導(dǎo)免疫耐受,防止自發(fā)性自身免疫糖尿病的發(fā)生。為使胰島素蛋白更有效地被腸相關(guān)淋巴組織吸收,本研究分別構(gòu)建了強(qiáng)免疫佐劑霍亂毒素B亞單位(cholera toxin B subunit, CTB)與胰島素原(proinsulin, PROIN)和胰島素B鏈(insulin B chian, INSB)的融合基因CTB-PROIN、CTB-INSB,探索了利用馬鈴薯生產(chǎn)1型糖尿病口服疫苗的可能性,主要研究?jī)?nèi)容和結(jié)果如下: 1.首先將融合基因CTB-PROIN、CTB-INSB克隆到大腸桿菌表達(dá)載體pET-30a(+)中,并誘導(dǎo)其表達(dá)。大腸桿菌表達(dá)的融合蛋白CTB-PROIN和CTB-INSB可與GM1糖苷脂結(jié)合,并具備CTB和胰島素的雙重抗原性。 2.在此基礎(chǔ)上,分別構(gòu)建了含有土豆塊莖特異性啟動(dòng)子Patatin、融合基因、馬鈴薯蛋白酶抑制劑基因Ⅱ(Pin2)的3’末端非翻譯區(qū)(UTR)序列和NOS終止子的兩個(gè)植物轉(zhuǎn)化載體,通過農(nóng)桿菌介導(dǎo)法轉(zhuǎn)化馬鈴薯,并獲得含有融合基因的轉(zhuǎn)基因馬鈴薯植株。轉(zhuǎn)基因植物表達(dá)的融合蛋白具有霍亂毒素和胰島素的雙重抗原性,可于神經(jīng)節(jié)苷脂GM1結(jié)合,具有潛在的生物活性。這為利用轉(zhuǎn)基因馬鈴薯免疫NOD小鼠,進(jìn)一步分析其免疫特性奠定了基礎(chǔ)。
[Abstract]:Oral islet specific autoantigen can induce immune tolerance and prevent spontaneous autoimmune diabetes mellitus. In order to make insulin protein more effectively absorbed by intestinal associated lymphoid tissue. In this study, cholera toxin B subunits (CTBs) and proinsulin were constructed respectively. The fusion gene CTB-PROIN-CTB-INSB of proin and insulin B strand insulin B chian (INSBs). The possibility of using potato to produce oral vaccine for type 1 diabetes mellitus was explored. The main contents and results were as follows: 1. Firstly, the fusion gene CTB-PROINOCTB-INSB was cloned into E. coli expression vector pET-30a (). The fusion proteins CTB-PROIN and CTB-INSB expressed by Escherichia coli can bind to GM1 glycosides and possess the double antigenicity of CTB and insulin. 2. On this basis, the potato tuber specific promoter Patatin was constructed, and the fusion gene was constructed. Potato protease inhibitor gene 鈪，

本文編號(hào)：1468857