摘要
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表述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。
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-crelacZ 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.
作者
顾晓雯
孙瑞林
费俭
GU Xiao-wen SUN Rei-lin FEI Jian(School of Life Science and Technology, Tongji University, Shanghai 200092, China Shanghai Biomodel Organism Science ＆ Technology Development Co.,Ltd., Shanghai 201203, China)
出处
《实验动物与比较医学》
CAS
2017年第2期89-93,共5页
Laboratory Animal and Comparative Medicine
基金
国家重点基础研究发展计划(2010CB945501)
