肝细胞Foxa2基因剔除小鼠的制备及鉴定

Generation and confirmation of Foxa2 gene knockout in hepatocyte in mice

目的制备肝细胞Foxa2基因特异性剔除的小鼠模型,并进行鉴定。方法采用CreloxP重组系统,在Cre重组酶的作用下,将小鼠肝细胞DNA上的Foxa2基因进行条件性的靶向剔除;并通过PCR和免疫荧光染色法进行鉴定。结果PCR在剔除型小鼠中未扩增出Foxa2基因的条带;同时定量分析也未测出其mRNA的表达;免疫荧光显示剔除型小鼠的Foxa2蛋白也为阴性。结论条件性基因剔除技术能成功地制备肝细胞Foxa2基因特异性剔除的小鼠模型。

s:Objective To generate Foxa2 loxP/loxPAFP.Cre mice and confirm the genetype.Methods Cre-loxP system was introduced to mediate targeting of the Foxa2 gene and to achieve hepatocyte specific expression of Cre; real-time PCR and immunofluorescence staining were used to confirm absence of Foxa2 in livers from Foxa2 loxP/loxPAFP.Cre mice.Results There is no band for Foxa2 by PCR, no Foxa2 mRNA expression by real-time PCR in Foxa2 loxP/loxPAFP.Cre mice.Conclusion Foxa2 is deleted efficiently in Foxa2 loxP/loxPAFP.Cre mice by conditional gene knockout.