LRP16肝特异性基因敲除小鼠模型的构建与鉴定

摘要

Objective To establish liver-specific leukemia-related protein 16 (LRP16) gene knockout mice model,and investigate the relationship between LRP16 gene and insulin resistance.Methods The knockout vector targeting exon 5 to exon 8 of LRP16 gene was constructed by molecular cloning and transfected to embryonic stem (ES) cell by electroporation.The selected positive ES cells was microinjected into the blastula of C57BL/6J mice.And the chimerical mice were born after transplantation of blastula into host mice.The LRP16-Loxp mice were born by crossing between chimerical mice and C57BL/6J mice,and the liver-specific LRP16 knockout mice were obtained by crossing between LRP16-Loxp mice and Alb-Cre mice.Results Eight positive ES cell cloning strains were obtained,and 2 chimerical mice,1 LRP16-Loxp mouse and 2 liver-specific LRP16 knockout mice were obtained to meet the research demand.Conclusion Liver-specific LRP16 knockout mice model is successfully established,which will be helpful in studying the correlation between LRP16 and insulin resistance.%目的 建立白血病相关蛋白16(leukemia-related protein 16,LRP16)肝脏特异性基因敲除小鼠模型,进而在体内研究LRP16基因与胰岛素抵抗的关系.方法 构建LRP16基因重组载体,通过电转打靶转染胚胎干细胞(ES细胞),将筛选出的阳性ES细胞克隆,显微注射至超排小鼠的囊胚腔后移植于受体小鼠,从子代中筛选嵌合体小鼠,与野生型C57小鼠杂交后得到LRP16-Loxp转基因小鼠,再与Alb-Cre小鼠杂交后得到LRP16肝特异性基因敲除小鼠并进行鉴定.结果 得到8株阳性ES细胞克隆,经显微注射后得到2只雄性嵌合体小鼠,与野生小鼠杂交后得到1只LRP16-Loxp转基因小鼠,与Alb-Cre小鼠杂交后再自交筛选出2只小鼠经鉴定为LRP16肝特异性基因敲除小鼠.结论 成功建立LRP16肝特异性基因敲除小鼠模型,为研究LRP16基因与胰岛素抵抗的关系奠定基础.