本文選題：熱帶螨 + 重組Blo�。� 參考：《中國(guó)畜牧獸醫(yī)》2017年07期

【摘要】：為了建立熱帶螨重組Blo t 5(rBlo t 5)變應(yīng)原誘導(dǎo)小鼠過(guò)敏模型,試驗(yàn)對(duì)熱帶螨Blo t 5基因序列進(jìn)行優(yōu)化與全基因合成,構(gòu)建了原核表達(dá)載體PQE80L-Blo t 5,并在大腸桿菌中進(jìn)行表達(dá)。利用鎳柱親和層析法純化了rBlo t 5蛋白,Western blotting結(jié)果顯示,純化的rBlo t 5蛋白能與Blo t 5鼠單克隆抗體結(jié)合。隨后將12只BALB/c小鼠隨機(jī)均分為對(duì)照組(PBS組)與誘導(dǎo)組(rBlo t 5組),通過(guò)腹腔注射致敏與氣道誘導(dǎo)后,酶聯(lián)免疫吸附試驗(yàn)(ELISA)檢測(cè)血清總IgE水平;鼠尾靜脈注射變應(yīng)原誘發(fā)應(yīng)激性過(guò)敏反應(yīng);HE染色觀察小鼠肺部炎癥嚴(yán)重程度。誘導(dǎo)小鼠過(guò)敏后發(fā)現(xiàn),與PBS組相比,rBlo t 5組小鼠血清總IgE水平顯著升高(P0.05),應(yīng)激性過(guò)敏反應(yīng)引起體溫顯著下降(P0.05),肺部組織細(xì)胞浸潤(rùn)顯著增加(P0.05)。本研究結(jié)果表明,原核表達(dá)純化的rBlo t 5蛋白能夠成功建立小鼠過(guò)敏模型。
[Abstract]:In order to establish a mouse allergic model induced by recombinant Blo t 5 (rBlo t 5), a prokaryotic expression vector PQE80L-Blo t 5 was constructed and expressed in Escherichia coli. The purified rBlo t 5 protein was purified by nickel column affinity chromatography. The results showed that the purified rBlo t 5 protein could bind to the monoclonal antibody of Blo t 5 mouse. Then 12 BALB / c mice were randomly divided into two groups: control group (PBS group) and induction group (rBlo t 5 group). The serum IgE levels were measured by enzyme-linked immunosorbent assay (Elisa) after intraperitoneal injection of sensitization and airway induction. The severity of lung inflammation in mice was observed by HE staining. Compared with PBS group, the serum total IgE level of mice induced by allergy was significantly higher than that of PBS group (P0.05), the body temperature of stress allergic reaction decreased significantly (P0.05), and the infiltration of lung tissue cells increased significantly (P0.05). The results showed that the purified rBlo t 5 protein could successfully establish mouse allergic model.
【作者單位】： 海南大學(xué)熱帶農(nóng)林學(xué)院動(dòng)物科技學(xué)院;海南大學(xué)熱帶農(nóng)林學(xué)院生物學(xué)院;
【基金】：國(guó)家自然科學(xué)基金青年項(xiàng)目(31402257) 海南省科技項(xiàng)目(ZDYF2017110) 海南省自然科學(xué)基金項(xiàng)目(20153084)
【分類號(hào)】：R-332;R392
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本文編號(hào)：2065578