Afp-cre-lacZ转基因小鼠的构建

目的 研究甲胎蛋白(Afp)阳性细胞的增殖分化在组织生长修复,肿瘤发生过程中的作用,建立Afp-CreERT转基因小鼠细胞示踪系统.方法 采用DNA雄原核显微注射方法获得Afp-CreERT转基因小鼠,筛选出合适的品系后,使之与Rosa26-lacZ工具小鼠杂交,获得Cre/lacZ双阳性的Afp-cre-lacZ转基因小鼠.结果 显微注射后,共出生小鼠56只,经PCR鉴定Cre阳性小鼠共4只,阳性小鼠传代后各为一系.筛选内源性Afp表达与Cre表达相对符合的品系作为Afp-CreERT转基因小鼠品系建系.Afp-CreERT转基因小鼠与Rosa26-lacZ工具小鼠杂交,经PCR鉴定后获得Cre/lacZ双阳性的Afp-cre-lacZ转基因小鼠.经实时PCR,X-gal染色,免疫荧光染色鉴定后得到Afp-cre-lacZ转基因小鼠细胞示踪系统,同时证实该系统能够正确示踪Afp表达阳性的细胞,同时也能够应用于肝损伤小鼠模型中.结论 成功构建了Afp-cre-lacZ转基因小鼠细胞示踪系统,为研究这类细胞的谱系发生提供了工具.

Construction of Afp-cre-lacZ Transgenic Mouse Model

Objective To study the function of alpha-fetoprotein (Afp) positive cells during tissues growth and repair or in tumorigenesis,Afp-cre-lacZ linage tracing mouse model was established.Methods Afp-CreERT mice were established by microinjection and identified by real time PCR.Afp-CreERT mice were crossed with Rosa26-1acZ mice.Cre/lacZ both positive mice were kept as Afp-cre-lacZ mice.Results Fifty-six transgenic mice were born after microinjection.Four of them genotyped Cre positive were kept as founder.Each founder was established a strain.Cre expression of each strain was identified via real time PCR.Thus Afp-CreERT transgenic mouse was established.The Cre/lacZ both positive offspring of Afp-CreERT mice were crossed with Rosa26-1acZ mice were kept as Afp-cre-lacZ mice.Identified by real time PCR,X-gal staining and immunofluorescence,Afp-cre-lacZ mice were observed that they could be used in lineage tracing and further studies.Conclusions The Afp-cre-lacZ mice established may be used as a tool for lineage tracing studies.