论文部分内容阅读
探讨自身抗原SSA/Ro-52kD 的抗原优势表位,为疾病机制的研究提供依据。根据计算机软件进行的蛋白质序列结构分析,采用PCR 法克隆自身抗原SSA/Ro-52kD 多肽片段的cDNA, 定向插入表达载体PGEX5X-1, 并且导入大肠杆菌中表达重组融合蛋白,用GST 亲和层析柱进行纯化,经免疫印迹法与病人阳性血清进行反应。结果表明片段Ro523 具有较强的抗原性。这提示SSA/Ro-52kD 的抗原优势表位主要存在于170~270 位。
To explore the antigen epitopes of autoantigens SSA / Ro-52kD, provide the basis for the study of disease mechanism. According to the analysis of protein sequence structure by computer software, the cDNA of self-antigen SSA / Ro-52kD fragment was cloned by PCR and inserted into expression vector PGEX5X-1. The recombinant fusion protein was introduced into E. coli and expressed by GST affinity chromatography The column was purified by Western blot with the patient’s positive serum reaction. The results showed that fragment Ro523 has strong antigenicity. This suggests that the epitope of SSA / Ro-52kD predominates at 170-270.