Afp-cre-lacZ转基因小鼠的构建

doi:10.3969/j.issn.1674-5817.2017.02.002

实验动物与比较医学 ›› 2017, Vol. 37 ›› Issue (2): 89-93.DOI: 10.3969/j.issn.1674-5817.2017.02.002

所属专题：实验动物资源开发与利用

Afp-cre-lacZ转基因小鼠的构建

顾晓雯¹, 孙瑞林², 费俭¹

1.同济大学生命科学与技术学院, 上海 200092;
2.上海南方模式生物科技股份有限公司, 上海 201203

收稿日期:2017-02-05 出版日期:2017-04-25 发布日期:2017-04-25
作者简介:顾晓雯(1988-),女,博士,从事生物化学与分子生物学研究。E-mail:17guxiaowen@tongji.edu.cn
基金资助:
国家重点基础研究发展计划(2010CB945501)

Construction of Afp-cre-lacZ Transgenic Mouse Model

GU Xiao-wen¹, SUN Rei-lin², FEI Jian¹

1. School of Life Science and Technology, Tongji University, Shanghai 200092, China;
2. Shanghai Biomodel Organism Science & Technology Development Co.,Ltd., Shanghai 201203, China

Received:2017-02-05 Published:2017-04-25 Online:2017-04-25

摘要/Abstract

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。

关键词: 甲胎蛋白(Afp), 转基因小鼠模型, 细胞示踪

Abstract: 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-lacZ 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-lacZ 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.

Key words: Alpha-fetoprotein (Afp), Transgenic mouse model, Lineage tracing

中图分类号:

Q95-33

顾晓雯,孙瑞林,费俭. Afp-cre-lacZ转基因小鼠的构建[J]. 实验动物与比较医学, 2017, 37(2): 89-93. DOI: 10.3969/j.issn.1674-5817.2017.02.002.

GU Xiao-wen,SUN Rei-lin,FEI Jian. Construction of Afp-cre-lacZ Transgenic Mouse Model[J]. Laboratory Animal and Comparative Medicine, 2017, 37(2): 89-93. DOI: 10.3969/j.issn.1674-5817.2017.02.002.

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Afp-cre-lacZ转基因小鼠的构建

Construction of Afp-cre-lacZ Transgenic Mouse Model

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