携带人衰变加速因子基因转基因小鼠的建立

Establishment of transgenic mice carrying human decay accelerating factor

目的 通过建立转人衰变加速因子 (DAF)基因小鼠 ,为研究异种器官移植的超急性排斥反应提供有效的研究手段。方法 采用受精卵显微注射技术 ,将人DAF基因导入小白鼠受精卵的原核中。Dot blot及Southern blot杂交确定阳性转基因小鼠。以Northern杂交法检测人DAF基因的表达情况。结果 基因注射后共生出小鼠 2 4只 ,4只为转基因小鼠 ,人DAF基因在其中 1只小鼠体内得到表达。结论 所导入的人DAF基因在小鼠的基因组中实现整合及表达。

Objective To develop transgenic mice carrying human decay accelerating factor(DAF)to provide an effective method for the study on xenogeneic hyperacute organ rejection. Methods A DNA fragment was isolated from plasmid, pSFFV DAF, consisting of SFFV 5'LTR as the promoter, hDAF cDNA and SV40 splice/poly A. Then the fragment was microinjected into male pronucleus of fertilized eggs of the Kunming mice. The genomic DNA was isolated from the the tail of the newborn mice with 4 week old. Dot blot and Southern blot hybridization methods were applied to identify the founder transgenic mice. Total RNA was isolated from kidney of transgenic mice, then Northern blot hybridization was used to detect the expression of hDAF gene in the transgenic mice. Results After microinjection of the gene, a total of 500 live fertilized eggs were obtained and were implanted into the oviducts of 24 pseudopregnant female mice. Human DAF gene was expression in one of the 4 transgenic mice. Conclusion The introduced hDAF gene has been integrated and expressed in those mice genome.