表达人自身抗原SSBLa重组体的构建及其鉴定（The construction and identification of the self antigen SSBLa recombinant body）.docVIP

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表达人自身抗原SSBLa重组体的构建及其鉴定（The construction and identification of the self antigen SSBLa recombinant body）.doc

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表达人自身抗原SSBLa重组体的构建及其鉴定（The construction and identification of the self antigen SSBLa recombinant body）

表达人自身抗原SSBLa重组体的构建及其鉴定（The construction and identification of the self antigen SSBLa recombinant body） The construction and identification of self antigen SSB/La recombinant body 10, 2010-11-11, deng ammei of the former kong xiantao One of the hallmarks of systemic autoimmune disease is the presence of some anti-nucleoprotein antibodies in circulation. Most autoantibodies, except antidna antibodies, have no direct effect on the pathogenesis of autoimmune diseases. However, specific autoantibodies are highly correlated with the corresponding autoimmune disease. The role of antigenic antibodies in autoimmune diseases helps us to understand the pathogenesis of autoimmune diseases and lay the foundation for early diagnosis and treatment of the disease. Autoantigen SSB/La can be sjogrens syndrome (SS), systemic lupus erythematosus (SLE) patients with autoimmune diseases such as serum recognition [1], but the antigen epitope of accurate positioning, table analysis of correlation between disease and is divided [2, 3]. The purpose of this project is to construct a recombinant body that expresses the antigenic SSB/La and to further express the recombinant antigen in e. coli, which is the basis for further study of the pathogenesis of the disease. 1 materials and methods 1 plasmid, strain, enzyme, and reagent: PGEX - 2T plasmid; JM 109 strain; AMV reverse transcriptase; Taq DNA polymerase, EcoR Ⅰ enzyme, BamH Ⅰ enzyme, T4DNA ligase (Pharmacia); QIAprep Spin Plasmid M iniprep Kit DNA. 1.2 the cDNA of SSB/La was cloned by reverse transcripting-pcr method: the EMBL library was retrieved, and a pair of primers designed for specific amplification SSB/La were designed by [4], the main standard of primers. The five prime of the primer added the appropriate sequence of endases to make the cloning gene fragment and the expression plasmid vector PGEX - 2T orienting and to ensure the correct reading frame. The primer was synthesized and purified by the Chinese academy of sciences. The 3