【摘要】： 由Ptpn22基因編碼的蛋白質(zhì)酪氨酸磷酸酶LYP在T細(xì)胞信號通路等信號轉(zhuǎn)導(dǎo)過程中起到重要的負(fù)調(diào)節(jié)作用,其功能異�？赡軙䦟�(dǎo)致機(jī)體免疫耐受的缺失,從而引起自身免疫疾病。本研究利用等位基因特異性PCR的方法,檢測中國人群自身免疫疾病患者體內(nèi)Ptpn22上C1858T單核苷酸多態(tài)性,并得出與以往相關(guān)研究不同的結(jié)論,即1858T等位基因并不能增加中國人群自身免疫疾病的患病幾率。另外,為了進(jìn)一步研究LYP的生理生化功能,本研究克隆了編碼LYP催化結(jié)構(gòu)域的Lyp-Cat,使其在E. coli Rosetta中高效表達(dá),并通過親和層析、制備電泳等技術(shù),提取純化了兩種LYP融合蛋白,GST-LYP-Cat和MBP-LYP-Cat。最后,本論文利用GST-LYP-Cat作為免疫抗原對小鼠進(jìn)行免疫,獲得了抗LYP-Cat的抗體,其靈敏度達(dá)到10-2ng,并利用正、負(fù)親和層析的方法對抗體進(jìn)行了純化,純化后的抗體可以有效識別Jurkat細(xì)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，

本文編號：2479454