携带人衰变加速因子基因转基因小鼠的建立

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杂交确定阳性围基因小鼠。以Ｎｏｒｈｅｒｎ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２４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ＤＡ

Establishment of transgenic mice carrying human decay accelerating factor

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.