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利用受精卵原核的显微注射技术 ,将人类HLA Ⅱ类DRα及DRB1 0 4 0 1两种基因 ,显微注射至C5 7BL 6×DBA 1杂交小鼠受精卵中 ;并移植至假孕受体鼠的输卵管内。实验先后有 8只鼠妊娠 ,稳定遗传五代 ,经PCR检测 95只HLA DRα、DRB1 0 4 0 1阳性。α 32P dCTP斑点杂交与Southern印迹杂交鉴定出 6 8只整合含有DRα及DRB1 0 4 0 1混合型的转基因小鼠 ,有效率为 2 0 .9%。经Northern杂交[1] 和RT PCR检测 ,其HLA DRα、DRB1 0 4 0 1基因在脾脏和肾脏中均有表达。结果说明 ,携带人类HLA Ⅱ类DRα和DRB1 0 4 0 1基因的转基因动物模型构建成功。
Using microinjection technique of prokaryotic nucleus, human HLA class II DRα and DRB10401 genes were microinjected into the fertilized eggs of C5 7BL 6 × DBA 1 hybrid mice and transplanted into pseudopregnant recipient Rat’s fallopian tube. The experiment has 8 pregnant rats, stable genetic five generations, by PCR detection of 95 HLA DRα, DRB1 0 4 0 1 positive. Sixty-eight transgenic mice that contained a mixture of DRα and DRB10401 were identified with α 32P dCTP dot blot hybridization and Southern blotting, with an efficiency of 20.9%. The results of Northern blot [1] and RT-PCR showed that HLA DRα and DRB10401 genes were expressed in spleen and kidney. The results showed that the transgenic animal model carrying human HLA class II DRα and DRB10401 gene was successfully constructed.