中文摘要：

目的将构建的CD59-linker-CD2融合基因真核表达载体在Jurkat细胞中稳定表达,研究CD59与CD2分子在T细胞信号转导中的相互作用,探讨CD59向T细胞内传递信号的相关机制。方法酶切鉴定构建的pIRES-CD59-linker-CD2融合基因真核表达载体,脂质体法转染Jurkat细胞,G418筛选建立稳定转染细胞系,免疫酶法检测转染细胞表面CD59的表达,Western blot检测转染细胞中CD59蛋白的表达;用CD59单克隆抗体作用于各组Jurkat细胞,使用激光共聚焦显微镜检测细胞浆内的Ca2＋,ELISA检测细胞上清中白介素2（IL-2）的变化,比较各组差异。结果经酶切鉴定证实携带CD59-linker-CD2融合基因的重组质粒扩增成功;免疫酶法和Western blot证实CD59在转染细胞中稳定表达,且转染组L-Jurkat细胞比正常组Ju-rkat细胞CD59表达量增加,二者比较其差别有统计学意义（P〈0.05）;与正常细胞相比,胞浆内Ca2＋和IL-2在转染细胞中均高表达,两组细胞比较其差别有统计学意义（P〈0.05）。结论 pIRES-CD59-linker-CD2融合基因真核表达载体可在Jurkat细胞中稳定表达,CD59mAb交联刺激后,CD59可通过CD2向细胞内传递信号,引起细胞内信号分子的一系列变化,为研究CD59与CD2在T细胞信号转导中的协同作用及进一步探讨CD59向T细胞内传递信号的机制奠定基础。

英文摘要：

The fusion expression vector of CD59 gene and CD2 gene was steadily expressed,which is aimed to study the reciprocity of CD59 and CD2 in T cell signal transduction and investigate CD59 transmitting signals to T cell.The eukaryotic expression vector containing CD59-linker-CD2 fusion gene was identified by digestion and transfected into Jurkat cells by liposome,while steady cells cloned could get through G418 screening.CD59 gene was detected by immunoenzyme method and Western blot.After CD59mAb cross-linking,the kinetic changes of i in T cell endochylema was identified by laser scanning confocal microscope,while interleukin 2（IL-2） in cell supernatants fluid was detected by ELISA.The eukaryotic expression vector was cloned correctly,which was confirmed by restriction endonuclease digestion.The results of immunoenzyme method and Western blot indicated that CD59 gene was steadily and highly expressed in transfected Jurkat cells compared with normal Jurkat cells,while the differences were statistically significant（P0.05）.i and IL-2 were highly expressed in transfected Jurkat cells compared with normal Jurkat cells,the differences were statistically significant（P0.05）.The eukaryotic expression vector containing CD59-linker-CD2 fusion gene could be steadily expressed in Jurkat cells.After CD59mAb cross-linking,CD59 could trigger a signaling response by means of CD2,which causes a series of changes of cell signal molecules.This results could be a foundation for the study of the synergistic effect of CD59 and CD2 in T cell signal transduction and CD59 transmitting signals to T cell.