转hDAF基因小鼠的构建(英文)  

转hDAF基因小鼠的构建

作　　者：王剑鹏[1] 王为先[2] 等 

机构地区：[1]天津泌尿外科研究所，中国天津300211 [2]中国科学院发育生物学研究所，中国北京100080

出　　处：《Developmental and Reproductive Biology》2000年第2期37-42,共6页发育与生殖生物学报（英文版）

摘　　要：Production of transgenic mice with human decay accelerating factor (hDAF) gene will provide an effective way to study how to overcome xenogenic hyperacute organ. The expression vector of hDAF gene, PSP 64HD, has been constructed. The PSP 64 HD DNA were lineared and microinjected into the fertilized eggs of mice by using male pronucleus microinjection technique. The transgenic eggs were transplanted into the uterus mice. When newborn mice were 4 weeks old, their tail DNAs were isolated to do Dot and Southern blot analysis for their-self identification. The results show that the constructed foreign gene has been integrated into the mouse genome. It means that the expression vector of hDAF gene and its transgenic mice have been constructed successfully in our experiments. For the transgenic mice, 77.9% of injected eggs survived after microinjection, and 3.4% in pups after development. The integration rate for hDAF gene is 10.5%.本工作构建了含有hDAF基因的转基因小鼠，以便研究hDAF基因能否消除异种器官移植中的排斥反应。采用DNA重组的方法构建hDAF基因的表达载体pSP64HP（Fig.1)。通过受精卵显微注射，将其中的目的的基因片段，转移到小鼠体内，建立转基因小鼠。再通过Dot blotting和Southern blotting杂交方法对出生小鼠的基因组特征进行查证。连续两次对直接裂解菌液做PCR扩增，筛选出重组质粒（Fig.2&3)，酶切图谱（Fig.4)和Southern杂交（Fig.5)分析结果与预期吻合，出现预期条带；小鼠受精卵注射后存活比率为77．9％，受精卵的发育率为3．4％，出生小鼠中，10．5％出现清晰杂交信号。表明：hDAF基因表达载体构建成功；并整合入小鼠基因组中。

关 键 词：hDNF基因 载体 转基因小鼠 移植免疫 

分 类 号：Q953.3[生物学—动物学]