小鼠干扰素limitin蛋白的表达及其鉴定

Expression and identification of murine interferon limitin protein

干扰素（IFN）作为机体抗病毒感染的第一道防线,已广泛应用于许多疾病的治疗中。作为Ⅰ型干扰素家族的一员,干扰素limitin于2000年被发现,然而目前国内外对该干扰素的研究尚停留在初级阶段。为了进一步研究limitin干扰素的生物学特性及其潜在的应用价值,本试验将密码子优化过的小鼠limitin基因克隆至载体pET-30a（＋）中,IPTG诱导表达limitin重组蛋白,用Ni-NTA亲和层析柱纯化,并用Westernblot进行验证。检测结果显示,表达的limitin重组蛋白的大小为23.86 ku,主要以包涵体形式存在,经纯化后获得高纯度的蛋白,并能被抗His标签的单克隆抗体特异性识别,最后利用生物信息学技术对这种干扰素蛋白进行了生物学预测与分析。结果表明,本研究成功表达并纯化出小鼠limitin重组蛋白。

Interferon consist the first line against viral infection and it has been widely used in the treatment of many diseases.As a member of theⅠtype interferon,interferon limitin was discovered in 2000.However,up to now,its studies which carried out in China and other countries are still in the primary stage.In order to further study its biological function of murine interferon limitin and promote its application in future,in this study,we initially cloned codon-optimized limitin gene into the vector pET-30 a（＋）,subsequently,the recombinant limitin protein was induced by IPTG,purified by NiNTA affinity chromatography and identified with Western-blot.The results showed that the recombinant limitin protein was 23.86 ku in molecular weight,and mainly expressed as inclusion bodies.It could specifically react with anti-His monoclonal antibody.Finally,we used bioinformatics technologies to predict and analyze the interferon protein.Overall,The murine IFN-limitin protein was successfully expressed and purified,which lays a foundation for the next research.