包含UI的人衰变加速因子重组基因在转基因鼠高效表达的研究  

Transgenic mice high-performancely expressing hDAF gene for xenotransplantation

作　　者：张玥[1] 王广有[1] 李胜芝[1] 马腾骧[1] 

机构地区：[1]天津市泌尿外科研究所,300211

出　　处：《中华实验外科杂志》2007年第8期999-1001,共3页Chinese Journal of Experimental Surgery

基　　金：天津市科技发展项目重点资助项目(043803411)

摘　　要：目的建立血管内皮细胞高效表达人衰变加速因子(hDAF)基因的转基因小鼠。方法采用受精卵显微注射法,将包含杂合内含子(UI)的人hDAF导入昆明白小鼠受精卵的雄原核内,然后将注射后仍健康的受精卵植入假孕母鼠输卵管内,待其自然分娩。应用聚合酶链反应(PCR)和Southern杂交检测G_0代小鼠hDAF基因的整合情况,应用流式细胞计数(FCM)检测小鼠外周血单核细胞(PBMCs)表面hDAF的表达,应用逆转录(RT)-PCR检测转基因小鼠心脏、肝脏、肾脏和肺脏人hDAF基因mRNA的表达。结果注射后卵的存活率和幼鼠出生率分别为83.8% (941/1123)和9.5%(89/941),外源基因的整合率为22.5%(20/89),FCM检测发现其中有7只转基因小鼠PBMCs有人hDAF表达,表达强度为人PBMCs的152%～243%,并且相应小鼠的心、肝、肾及肺组织均有人hDAF mRNA表达。结论UI可以使hDAF在转基因鼠各主要脏器高效表达。Objective To research high-performance expression of human decay-accelerating factor （hDAF） gene in transgenic mice for xenotransplantation. Methods Transgenic mice expressing hDAF were generated by microinjection of a recombinant hDAF gene containing UI enhancer under the control of the human ICAM-2 promoter. Offspring were tested for transgene integration by PCR and Southern-blotting analysis,and for hDAF expression in peripheral blood mononuclear cells （PBMCs） by flow cytometry （FCM） and RT-PCR was performed to detect the distribution of hDAF mRNA in the heart,liver, kidney and lung of transgenic mice. Results 941 injected and survived zygotes were implanted into the oviducts of pseudopregnant foster mothers. Integration rate of hDAF gene was 22.5% （20/89）. FCM analysis revealed hDAF was expressed in PBMCs of 7 transgenic miceand the expression intensity was 152% to 243% of that in human PBMCs. Expression of hDAF mRNA was detected in the heart, liver, kidney and lung of transgenic mice. Conclusion A transgenic mice model with overexpression of hDAF was successfully established for xenotransplantation.

关 键 词：异种移植 衰变加速因子 基因表达 

分 类 号：R346[医药卫生—基础医学]