凝血因子Ⅸ基因剔除小鼠近交系的建立

Establishing inbred strain of coagulation factor Ⅸ-knockout mouse

目的 :建立可稳定遗传、临床症状显著的凝血因子 (F )基因剔除小鼠近交系。 方法 :以 PCR扩增检测小鼠尾组织 DNA,一期法检定小鼠血浆 F 活性和血浆凝血酶原时间 (PT)、白陶土部分凝血活酶时间 (KPTT)值 ;取以上结果为全阳性的小鼠以选优法进行全同胞兄妹交配 ,每代选用第一胎小鼠留种 ,第 8代开始筛选纯合子。 结果 :F8代 F 基因剔除小鼠PCR检测阳性率 92 % ,到 F1 2 代小鼠阳性率达 10 0 % ;F 活性 (2 .4 7± 1.75 ) %较正常小鼠 (14 8.18± 4 6 .98) %明显降低 (P<0 .0 1) ;F 基因剔除小鼠已近交培育到第 12代 ,遗传性状稳定 ,并有自发出血倾向。结论 :成功建立了纯合子 F 基因剔除小鼠近交系 ,该小鼠符合人血友病

Objective:To establish a inbred strain of the coagulation factor Ⅸ knockout mice with stable heredity and obvious clinical symptoms. Methods:The genomic DNA from tail tissue of mice were analyzed by PCR. F Ⅸ activity plasma prothrombin time and kadin partial thromboplastin time were measured by one stage method.The positive mice were mated with brother sister by optimum seeking method.The stocks of every generation were selected from first born mice.The homogeneous mice were selected after F 8. Results: PCR analysis of F Ⅸ mice(F 8) was 92% positive and F Ⅸ mice(F 12 ) was 100% positive. F Ⅸ activity (2.47±1.75)% was lower than normal mice (148.18±46.98)%. F Ⅸ knockout mice were developed 12th generation and there were spontaneous bleeding in the factor Ⅸ knockout mice. Conclusion:The result showed that the coagulation F Ⅸ knockout mice inbred strain were established and exhibited a phenotype similar to human hemophilia B.