人sCD40L转基因小鼠模型的构建

目的研究阻断CD40-CD40L共刺激信号通路对移植皮肤免疫排斥反应的影响。方法通过RT-PCR技术克隆了呈可溶性表达的CD40L分子胞外区(sCD40L),利用K14启动子构建了sCD40L皮肤特异性表达载体,并利用该载体制备了转基因小鼠。结果所克隆的CD40L胞外区片段其大小及序列符合预期;以哺乳动物表达载体PCI为骨架,通过DNA重组,获得了含K14启动子和sCD40L编码区的皮肤特异性表达载体K14-sCD40L;通过显微注射和胚胎移植,PCR筛选检测:49只G0代小鼠有1只小鼠扩增出特异性条带,阴性对照无条带。结论成功建立sCD40L转基因阳性小鼠。

Establishment of a human sCD40L transgenic mouse model

Objective To study the effects of blockade of CD40-CD40L pathway on the survival of xenogeneic skin graft.Methods The extracellular domain of human CD40 ligand（CD40L） was cloned by RT-PCR,and thereby a skin-specific transgene expression vector for the soluble CD40L molecule（sCD40L） was constructed by keratine 14（K14） promoter.Using the sCD40L skin-specific transgene construct,transgenic mice were generated by pronuclear microinjection.Results The sCD40L fragment with expected size and sequence was correctly cloned and its skin-specific expression vector correctly constructed.By microinjection,embryo transfer,and PCR screening test,one of 49 G0 mice had amplified specific bands,which was absent in the negative controls.Conclusion We have successfully established a sCD40L transgenic positive mouse model.