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. Key words: Transplantation,heterologous; Decay accelerating factor; Mouse,transgenic