本文選題：弓形蟲 + 表面抗原(SAG)�。� 參考：《福建農(nóng)林大學(xué)學(xué)報(bào)(自然科學(xué)版)》2017年03期

【摘要】：將沙門氏菌鞭毛素和弓形蟲免疫優(yōu)勢(shì)表面抗原1(SAG1)融合表達(dá),研究其在小鼠上激發(fā)的體液免疫應(yīng)答.首先,采用PCR擴(kuò)增SAG1片段.其次,將SAG1連接到原核表達(dá)載體pET-28a中構(gòu)建表達(dá)質(zhì)粒pET-28a-SAG1;連接到實(shí)驗(yàn)室已有的pET-28a鞭毛素質(zhì)粒中構(gòu)建表達(dá)質(zhì)粒pET-28a-F-SAG1;質(zhì)粒轉(zhuǎn)化至大腸桿菌BL21(DE3)菌株中并進(jìn)行表達(dá).將純化后的蛋白免疫小鼠,每隔2周免疫一次,共免疫3次,第3次免疫后15 d取小鼠血清.最后,采用蛋白印跡檢測(cè)蛋白的大小;用酶聯(lián)免疫吸附方法檢測(cè)血清中多克隆抗體的效價(jià).結(jié)果表明,誘導(dǎo)出的SAG1蛋白大小為30 ku,F-SAG1大小為70 ku.酶聯(lián)免疫吸附方法檢測(cè)的D450 nm均為陰性對(duì)照的兩倍,表明該多克隆抗體具有較高效價(jià).可見(jiàn),鞭毛素可以作為分子佐劑促進(jìn)弓形蟲亞單位疫苗的體液免疫應(yīng)答.
[Abstract]:Salmonella flagellin was fused with Toxoplasma gondii immune dominant surface antigen 1 SAG1 to study its humoral immune response in mice. First, the SAG1 fragment was amplified by PCR. Secondly, the expression plasmid pET-28a-SAG1 was constructed by ligating SAG1 into the prokaryotic expression vector pET-28a, and the expression plasmid pET-28a-F-SAG1 was constructed by ligating into the flagellum diathesis granules of pET-28a. Mice were immunized with purified protein once every 2 weeks for 3 times. The serum of mice was collected 15 days after the third immunization. Finally, the size of protein was detected by Western blotting, and the titer of polyclonal antibody in serum was detected by enzyme-linked immunosorbent assay (Elisa). The results showed that the induced SAG1 protein size was 30 kuo F-SAG 1 and 70 ku. The D450 nm detected by enzyme-linked immunosorbent assay (Elisa) was twice as high as that of the negative control, indicating that the polyclonal antibody had high titer. Therefore, flagellin can be used as a molecular adjuvant to promote humoral immune response of Toxoplasma gondii subunit vaccine.
【作者單位】： 福建農(nóng)林大學(xué)動(dòng)物科學(xué)學(xué)院/福建省動(dòng)物藥物工程實(shí)驗(yàn)室;
【基金】：國(guó)家自然基金青年項(xiàng)目(31502058) 福建省自然科學(xué)基金面上項(xiàng)目(2015J01075) 福建省中青年教師教育科研項(xiàng)目(JA15159) 大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計(jì)劃項(xiàng)目(201610389148、201510389029)
【分類號(hào)】：S852.7

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