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目的预测相对分子量为52000的Ro/SSA自身抗原的可能表位,并选择性克隆含亮氨酸拉链氨基酸序列的抗原片段。方法用蛋白结构分析软件对52000Ro/SSA抗原结构进行分析,从人心脏来源的cDNA中通过聚合酶链反应(PCR)扩增得到编码第119至264位氨基酸片段的cDNA,插入表达载体,转染大肠杆菌并表达融合蛋白。结果计算机分析显示该抗原中有α螺旋结构,整个分子抗原性强,柔韧性一般,表面可及性差。PCR产物长440bp,重组融合蛋白相对分子质量为28000,测序鉴定保证了研究的正确性。结论52000Ro/SSA多肽抗原性好,所获得的抗原片段为今后研究亮氨酸拉链氨基酸序列在该抗原表位形成中的作用奠定了基础。
Objective To predict the possible epitope of Ro / SSA autoantigen with relative molecular weight of 52000 and to selectively clone the antigenic fragment containing the amino acid sequence of leucine zipper. Methods The protein structure analysis software was used to analyze the structure of 52000Ro / SSA antigen. The cDNA encoding the amino acid fragment from the 119th to the 264th positions was amplified by polymerase chain reaction (PCR) from human cardiac cDNA, inserted into the expression vector, and transfected E. coli and express the fusion protein. Results Computer analysis showed that the antigen has an α-helix structure, the whole molecule has strong antigenicity, general flexibility and poor surface accessibility. The length of the PCR product was 440bp, and the relative molecular mass of the recombinant fusion protein was 28,000. Sequencing identification ensured the correctness of the study. Conclusion The antigenicity of 52000Ro / SSA polypeptide is good, and the antigenic fragment obtained lays the foundation for the future study on the function of leucine zipper amino acid sequence in the formation of this epitope.