一种重组口蹄疫疫苗的原核表达、纯化及其活性检测

来源 :细胞与分子免疫学杂志 | 被引量 : 0次 | 上传用户:zhenlijinping
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目的:利用原核表达系统诱导表达一种可以作为口蹄疫疫苗的重组蛋白J11-G,并对这种蛋白进行纯化以及活性检测。方法:对已经构建的表达载体PET28a-J11-G进行酶切鉴定,将鉴定后的重组载体PET28a-J11-G转化入大肠杆菌BL21,在BL21细菌细胞中诱导表达目的蛋白,用改良的镍亲和层析法纯化目的蛋白,以ELISA检测纯化所得蛋白的活性。结果:长期冻存的重组载体PET28a-J11-G结构完整,其在大肠杆菌BL21中经1mmol/LIPTG诱导后表达出相对分子质量(Mr)约为47000的目的蛋白,经纯化后获得了产率和纯度均很高的重组蛋白J11-G,ELISA检测证明该蛋白活性良好。结论:成功地对重组蛋白J11-G进行了诱导表达,改良法有利于蛋白产率和纯度的提高,纯化蛋白具有较好的免疫活性。重组蛋白J11-G将可能成为一种抗口蹄疫疫苗。 OBJECTIVE: To induce the expression of a recombinant J11-G protein that can be used as a foot-and-mouth disease vaccine by using prokaryotic expression system and to purify this protein and test its activity. Methods: The constructed expression vector PET28a-J11-G was identified by restriction enzyme digestion. The identified recombinant vector PET28a-J11-G was transformed into E.coli BL21 and expressed in BL21 cells. Purification of the target protein by chromatography, and the activity of the purified protein was detected by ELISA. Results: The recombinant vector PET28a-J11-G, which had been frozen for a long time, was structurally intact. After induced by 1mmol / L IPTG in E. coli BL21, the recombinant protein PET28a-J11-G was expressed with a molecular weight of 47,000 (Mr) And high purity of recombinant protein J11-G, ELISA test showed that the protein activity is good. Conclusion: The recombinant protein J11-G was induced successfully. The improved method was beneficial to the increase of protein yield and purity, and the purified protein had good immunocompetence. Recombinant protein J11-G will likely become an anti-foot-and-mouth disease vaccine.
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