中国免疫学杂志Issue(10)：1369-1373,5.DOI:10.3969/j.issn.1000-484X.2014.10.015
MuSK-mCherry融合荧光蛋白的构建及对重症肌无力患者MuSK抗体的检测
Construction of MuSK-mCherry fusion fluorescent protein and used for detection of MuSK antibodies in myasthenia gravis
摘要
Abstract
Objective:To construct a fusion protein of extracellular domain peptide fragment of muscle specific kinase ( MuSK) and fluorescent protein mCherry ,and used as antigen in the detection of antibodies against MuSK ( MuSKAb ) in the sera of patients with myasthenia gravis ( MG).Methods:The mCherry gene was amplified by PCR from vector pRSET-B and cloned into pGEM-T Easy Vector,and furthermore, cloned into Eukaryotic expression vector pMT /BiP/V5-His ( MuSK), which contains MuSK extracellular domain 22-452 amino acid peptide fragment gene to construct the fluorescent fusion protein gene MuSK -mCherry.The recombinant vector was subsequently transfected into drosophila S 2 cells for expression.The expressed fusion proteins were verified in confocal mi-croscope ,and used as antigen in the detection of MuSKAb in sera of MG patents in fluorescence immunoprecipitation test .Results:The fluorescent fusion protein MuSK-mCherry was successfully constructed and expressed.The MuSKAb in sera of patents with MG could be detected in fluorescence immunoprecipitation test using the constructed MuSK-mCherry fusion protein as antigen.Conclusion: It is available to use the constructed fluorescent fusion protein MuSK-mCherry as antigen in fluorescence immunoprecipitation test for the detection of MuSKAb in sera of patents with MG.关键词
重症肌无力/肌肉特异性激酶/荧光蛋白/肌肉特异性激酶-红色荧光蛋白融合蛋白Key words
Myasthenia gravis (MG)/Muscle specific kinase (MuSK)/Fluorescent protein/MuSK-mCherry fusion protein分类
医药卫生引用本文复制引用
金权鑫,许嘉珍,魏枫,李芳芳,王岩,李红花,金松竹,孟繁平..MuSK-mCherry融合荧光蛋白的构建及对重症肌无力患者MuSK抗体的检测[J].中国免疫学杂志,2014,(10):1369-1373,5.基金项目
国家自然科学基金项目（81160373，81360458）资助。 （）
