转hMCP/hCD_(59)双基因小鼠的构建、传代及功能测试

Foundation, Heredity and Function Test of Double Gene Mouse with hMCP and hCD_(59)

目的比较转染两个补体调节蛋白(CRP)基因与转染单一CRP基因对异种细胞的保护作用。方法采用双基因混合注射的方法,将人MCP(膜辅因子蛋白)和CD59(膜反应性溶解抑制物)两种基因导入521枚KM小鼠的受精卵原核。结果得到原代转hCD59基因小鼠14只,转hMCP基因小鼠12只,hCD59/hMCP双基因共整合的小鼠7只,转基因效率5%。通过5只G0代转双基因小鼠和非转基因小鼠的繁殖,得到F1转hMCP小鼠14只,转hCD59小鼠16只,转双基因小鼠10只,F1双基因占33.3%。用10%新鲜人血浆体外灌注小鼠离体心脏,转双基因组心脏搏动时间(138±25min)明显长于单基因组(hCD59组78±27min,hMCP组43±21min)和非转基因对照组(20±12min)。结论转染两个CRP基因比转染单一CRP基因对异种细胞有更好的保护作用。

Objective Comparing protective effect on the cell from one CRP or two CRPs. Method Double gene mouse was founded by mixed injection. Mixed human MCP and CD59 gene was injected into pronuclear of 521 mice zygote. Results In all of the newborn mouse, there are 14 with CD59 gene, 12 with MCP gene, 7 with double gene. Total transgene rate was 5.0 percent. Five double gene founders mated with nontransgene mouse, which their baby had 14 with MCP gene, 16 with CD59 gene, 10 with double gene corresponded to 33.3 percent of baby mouse. The hearts in vitro were taken out to be perfused with fresh human blood plasma, the average heart beating time was 138±25 min in the hCD59/hMCP group. The time is significantly longer than that of other group （78±27 min in the hCD59 group, 43±21 min in the hMCP group, and 20±12 min in the nontransgenic group）. Conclusion The organ of double gene mouse is better to resist immunity attacking than one gene mouse.