论文部分内容阅读
真菌免疫调节蛋白(Fip)是真菌中重要的药理成分之一。为进一步开发真菌中Fip资源,从云芝中克隆了云芝免疫调节蛋白基因Fip-cve,对Fip-cve的理化性质和生物信息学进行了研究,并将其转入酵母表达载体中,成功获得了酵母转化子。结果表明:Fip-cve基因扩增片段长为336 bp,该序列编码111个氨基酸;Fip-cve蛋白的二级结构是以无规卷曲为主,同时含有35.71%的延伸链,还含有12.35%的α-螺旋,主要位于N端;疏水性分析表明该基因编码的蛋白是亲水性蛋白,二级结构分析也解释了该蛋白的结构特征;SDSPAGE电泳结果显示Fip-cve目的蛋白均在酵母中获得了高效表达,目的蛋白分子量约为13 ku;生物信息学分析结果表明,Fip-cve与灵芝属真菌免疫调节蛋白具有很高的同源性,特别是与小孢灵芝具有更高的同源性。该研究初步建立了Fip-cve的真核表达体系,以期为Fip-cve的进一步开发提供参考。
Fungal immunomodulatory protein (Fip) is one of the important pharmacological components in fungi. In order to further develop the Fip resources in fungi, Fip-cve was cloned from Yunzhi, and the physicochemical properties and bioinformatics of Fip-cve were studied and transformed into yeast expression vector. Yeast transformants were obtained. The results showed that the Fip-cve gene was amplified with a length of 336 bp and encoded 111 amino acids. The secondary structure of Fip-cve protein was random coil with 35.71% extension chain and 12.35% The results of SDS-PAGE showed that the target proteins of Fip-cve were expressed in yeast The molecular weight of the target protein was about 13 ku. Bioinformatics analysis showed that Fip-cve had high homology with fungi immunoregulatory proteins of G. lucidum, Source. The research initially established the eukaryotic expression system of Fip-cve, in order to provide a reference for the further development of Fip-cve.