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目的 克隆人ｓＣＤ４０Ｌ基因片段，并在原核细胞中表达。方法用ＲＴ－ＰＣＲ技术，从激活的人外周血淋巴细胞总ＲＮＡ中，扩增ＣＤ４０Ｌ胞外区ｃＤＮＡ，并克隆至载体ｐＧＥＭ－Ｔ。测序验证后，转至载体ＰＱＥ３１中，并在大肠杆菌Ｍ１５中进行表达，最后通过亲和层析柱得到纯化蛋白。结果纯化所得的产物经Ｗｅｓｔｅｒｎ ｂｌｏｔ证实，确为人ｓＣＤ４０Ｌ蛋白。结论 应用基因重组法构建了人ｓＣＤ４０Ｌ基因片段，并在原核细胞内成功地进行了表达，为今后进一步研究ＣＤ４０Ｌ与凋亡、疾病的发病机制及临床治疗奠定了基础。
Objective To clone human sCD40L gene and express in prokaryotic cells. Methods The cDNA of CD40L extracellular region was amplified by RT-PCR from activated human peripheral blood lymphocytes and cloned into vector pGEM-T. After sequencing, the vector was transformed into vector PQE31 and expressed in E. coli M15. Finally, the purified protein was obtained by affinity chromatography. Results The purified product was confirmed by Western blot, indeed human sCD40L protein. Conclusion The human sCD40L gene fragment was constructed by gene recombination and successfully expressed in prokaryotic cells, which laid the foundation for further study of CD40L and apoptosis, pathogenesis and clinical treatment of disease.