异种肝移植模型-转人CD59蛋白基因小鼠的建立

Animal model for heterologous transplantation-generating of transgenic mice that expresses hCD59 cDNA

目的 通过显微注射法产生用于异种移植研究的表达人CD5 9蛋白的转基因小鼠模型。方法 实验中利用OMT作为目的基因hCD5 9cDNA的启动子 ,CMV作为报告基因GFP的启动子 ,构建重组质粒 pGOC。通过显微注射法 ,把CMV GFP OMT CD5 9基因片断注射入昆明白小鼠受精卵。结果 产生出 86只F0 代转基因小鼠。鼠尾基因组DNA经PCR法检测 ,12只阳性 ,进一步用Southernblot法检测 1只阳性。结论 通过显微注射法 ,外源基因hCD5 9cDNA在小鼠基因组中得到整合 ,产生出了阳性转人CD5 9蛋白基因小鼠 。

Objective To develop transgenic mouse model for heterologous transplantation study.Methods hCD59 cDNA was used as a targeting gene under the control of OMT (ovine metallothionein)promoter,and GFP (Green fluorescent protein) as a marker for gene expression under the control of CMV (cytomegalovirus)promoter. CMV-GFP-OMT-CD59 fragments were microinjected into fertilized mouse ova using microinjection.Results All of 86 transgenic Kunming mice was obtained by microinjection. PCR analysis showed 12 out of 86 mice (newborn mice) transgenic.Southern blot analysis showed 1 out of 86 transgenic. Conclusion hCD59 cDNA has integrated into mice genomic DNA. Transgenic mice have been obtained for heterologous transplantation study.