旋毛虫抗原基因T668的定点突变及原核表达

发布时间：2018-07-05 19:22

本文选题：旋毛虫 + T基因　；参考：《中国兽医学报》2017年04期

【摘要】：旋毛虫抗原基因T668是本实验室发现的1条高反应原性的特异性抗原基因,该基因编码旋毛虫丝氨酸蛋白酶。本试验利用重叠PCR的方法对旋毛虫抗原基因T668成功实现酶活性中心重要位点定点突变改造,将酶活性中心的结合位点259位亲水性丝氨酸和催化三联体中240位的丝氨酸分别突变为疏水性丙氨酸,并构建、纯化了可溶性表达的双位点突变的重组蛋白,后经Western blot分析表明所表达的重组蛋白能够与旋毛虫感染26d阳性血清发生特异性反应,具有良好的反应原性。本试验为后续开展对该丝氨酸蛋白酶的研究及应用奠定了坚实的基础。
[Abstract]:Trichinella spiralis antigen gene T668 is a highly reactive specific antigen gene which encodes serine protease of Trichinella spiralis. In this experiment, the gene T668 of Trichinella spiralis antigen was successfully transformed into a site-directed mutation at an important site of the enzyme active center by overlapping PCR. The binding site 259 of the enzyme active center and the 240 position serine of the catalytic triad were mutated to hydrophobic alanine, respectively, and the soluble recombinant protein with double site mutation was constructed and purified. Western blot analysis showed that the expressed recombinant protein could react specifically with the positive serum of Trichinella spiralis for 26 days. This experiment laid a solid foundation for the further research and application of the serine protease.
【作者单位】：吉林大学人兽共患病研究所;
【基金】：国家自然科学基金资助项目(31520103916)
【分类号】：S852.7

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