【摘要】 目的构建系统性红斑狼疮(systemiclupuserythematosus,SLE)外周血单个核细胞基因表达系列分析(serialanalysisofgeneexpression,SAGE)文库。方法从3例家族性SLE患者外周血单个核细胞分离PolyARNA,采用改良的SAGE技术构建SAGE文库,随机挑选阳性克隆进行菌落PCR鉴定及对部分阳性克隆进行测序。结果初步构建的SAGE文库已获得600个克隆,随机取50个进行菌落聚合酶链反应(PCR)分析,其中43个克隆中质粒插入片段大于300bp,占86%。部分克隆测序证实含有序列正确的SAGE标签。结论应用改良SAGE的技术成功构建了SLE外周血单个核细胞SAGE文库,为筛选SLE患者差异表达基因奠定了基础。更多还原

【Abstract】 Objective To construct serial analysis of gene expression (SAGE) library of peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus(SLE). Methods Poly A RNA extracted from PBMCs was used to construct SAGE library on family SLE patients by means of SAGE technique with some modifications. Randomly transformants were analyzed by colony PCR followed by sequencing. Results SAGE library of PBMCs of patients with SLE was set up with high quality. Eighty-six percent of colonies contained the expected insert fragment longer than 250 bp confirmed by colony PCR and subsequent sequencing. Conclusion SAGE library of PBMCs of patients with SLE is successfully constructed which set up a foundation for screening differentially expressed genes in patients with SLE.更多还原