CAJ | 학술논문

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基因导人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.结论所导入的人DAF基因在小鼠的基因组中实现整合及表达.
목적 통과건립전인쇠변가속인자(DAF)기인소서,위연구이충기관이식적초급성배척반응제공유효적연구수단.방법 채용수정란현미주사기술,장인DAF기인도인소백서수정란적원핵중.Dot-blot급Southern-blot잡교학정양성전기인소서.이Northern잡교법검측인DAF기인적표체정황.결과 기인주사후공생출소서24지,4지위전기인소서,인DAF기인재기중1지소서체내득도표체.결론 소도입적인DAF기인재소서적기인조중실현정합급표체.
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.