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人sCD40L转基因小鼠模型的构建
引用本文:艾必燕,樵星芳,周晓杨,王露露,杨扬,陈建康,刘勤. 人sCD40L转基因小鼠模型的构建[J]. 中国实验动物学报, 2012, 20(4): 35-40
作者姓名:艾必燕  樵星芳  周晓杨  王露露  杨扬  陈建康  刘勤
作者单位:[1]重庆市中药研究院,重庆400065 [2]第三军医大学,重庆400038
基金项目:国家“重大新药创制”科技重大专项(2010ZX09401-306-1-6);重庆市科技创新能力建设(CSTC,2010CB5013).
摘    要: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。

关 键 词:转基因小鼠  sCD40L  共刺激信号通路

Establishment of a human sCD40L transgenic mouse model
AI Bi-yan,QIAO Xing-fang,ZHOU Xiao-yang,Wang Lu-lu,YANG Yang,CHEN Jlan-kang,LIU Qin. Establishment of a human sCD40L transgenic mouse model[J]. Acta Laboratorium Animalis Scientia Sinica, 2012, 20(4): 35-40
Authors:AI Bi-yan  QIAO Xing-fang  ZHOU Xiao-yang  Wang Lu-lu  YANG Yang  CHEN Jlan-kang  LIU Qin
Affiliation:1. Chongqing Academy of Chinese Materia Medica, Chongqing 400065, China; 2. the Third Military Medical University, Chongqing 400038)
Abstract: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.
Keywords:Transgenic mice  sCD40L  Co-stimulatory pathway
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