Ptpn22 1858位单核苷酸多态性与中国人群自身免疫疾病相关性研究
发布时间：2019-05-17 22:31
【摘要】： 由Ptpn22基因编码的蛋白质酪氨酸磷酸酶LYP在T细胞信号通路等信号转导过程中起到重要的负调节作用,其功能异常可能会导致机体免疫耐受的缺失,从而引起自身免疫疾病。本研究利用等位基因特异性PCR的方法,检测中国人群自身免疫疾病患者体内Ptpn22上C1858T单核苷酸多态性,并得出与以往相关研究不同的结论,即1858T等位基因并不能增加中国人群自身免疫疾病的患病几率。另外,为了进一步研究LYP的生理生化功能,本研究克隆了编码LYP催化结构域的Lyp-Cat,使其在E. coli Rosetta中高效表达,并通过亲和层析、制备电泳等技术,提取纯化了两种LYP融合蛋白,GST-LYP-Cat和MBP-LYP-Cat。最后,本论文利用GST-LYP-Cat作为免疫抗原对小鼠进行免疫,获得了抗LYP-Cat的抗体,其灵敏度达到10-2ng,并利用正、负亲和层析的方法对抗体进行了纯化,纯化后的抗体可以有效识别Jurkat细胞中所含有的LYP。
[Abstract]:Protein tyrosine phosphatase LYP encoded by Ptpn22 gene plays an important negative role in signal transduction such as T cell signal pathway. Abnormal function may lead to the lack of immune tolerance, which may lead to autoimmune diseases. In this study, the C1858T single nucleotides polymorphism on Ptpn22 of patients with autoimmune diseases in Chinese population was detected by allele-specific PCR, and it was concluded that it was different from the previous studies. That is, 1858T allele did not increase the risk of autoimmune diseases in Chinese population. In addition, in order to further study the physiological and biochemical functions of LYP, the Lyp-Cat, encoding LYP catalytic domain was cloned to express it in E. coli Rosetta, and the Lyp-Cat, was prepared by affinity chromatography and electrophoresis. Two kinds of LYP fusion proteins, GST-LYP-Cat and MBP-LYP-Cat., were extracted and purified. Finally, in this paper, GST-LYP-Cat was used as immune antigen to immunize mice, and the antibody against LYP-Cat was obtained, the sensitivity of which reached 10 鈮，
本文编号：2479454
[Abstract]:Protein tyrosine phosphatase LYP encoded by Ptpn22 gene plays an important negative role in signal transduction such as T cell signal pathway. Abnormal function may lead to the lack of immune tolerance, which may lead to autoimmune diseases. In this study, the C1858T single nucleotides polymorphism on Ptpn22 of patients with autoimmune diseases in Chinese population was detected by allele-specific PCR, and it was concluded that it was different from the previous studies. That is, 1858T allele did not increase the risk of autoimmune diseases in Chinese population. In addition, in order to further study the physiological and biochemical functions of LYP, the Lyp-Cat, encoding LYP catalytic domain was cloned to express it in E. coli Rosetta, and the Lyp-Cat, was prepared by affinity chromatography and electrophoresis. Two kinds of LYP fusion proteins, GST-LYP-Cat and MBP-LYP-Cat., were extracted and purified. Finally, in this paper, GST-LYP-Cat was used as immune antigen to immunize mice, and the antibody against LYP-Cat was obtained, the sensitivity of which reached 10 鈮，
本文编号：2479454
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