摘要小鼠是哺乳动物研究实验中首选的一种模式生物。经过长时间的努力发展，培育转基因和基因打靶小鼠已经是很常见的事了，现在已经可以得到在核酸水平上的突变品系。目前，像这种基因组修饰变得越来越精确，而且在基因水平上标记不同的细胞群已变成非常普遍的研究[1]。88706

    人类基因组计划（HGP）的完成之后，生物学研究中急需一种有效的基因功能分析方法。而基因敲除小鼠模型的应用，为研究基因的功能提供了大力支持并可以研究基因在疾病发生中的作用，从而揭示疾病的发生机制，推动医药学研究的发展。基因捕获和基因打靶是两种不同的，通过胚胎干细胞（ES细胞）制作基因敲除小鼠的技术[2]。

    目前Cre-LoxP重组酶系统在小鼠基因打靶中被广泛应用，相比传统的转基因技术，应用该系统的基因敲除具有时空特异性，优化了转基因小鼠模型。应用Cre-LoxP重组酶系统构建的转基因小鼠可在小鼠局部实现基因敲除，很大程度上避免小鼠过早死亡，为后续研究提供可行性。

    本文通过基因敲入构建出Sox2CreER转基因小鼠，并经过扩大繁殖。然后利用PCR技术鉴定筛选出成功构建的Sox2CreER转基因小鼠，为后续的Cre-LoxP重组实验提供材料支持。

毕业论文关键词：转基因小鼠；PCR扩增；基因型鉴定

Abstract Mice as mammals preferred mode of biological genetic research。 Through the efforts of the past few decades, cultivating target genes and transgenic mice has become a very common work, with current technology can obtain mutant strains in the nucleic acid level。 Now, such as the genome modification is becoming more and more sophisticated, and mark at the gene level different cells has become a very common research。

As the human genome project (HGP) is completed, the gene era of biology is an urgent need to an effective method for gene function analysis。 Application of gene knockout mice model for the study of gene function and find new cures for human diseases provides strong support for the intervention measures。 Gene targeting and gene capture is two different by embryonic stem cells (ES cells) production technology of gene knockout mice。源Q于D优G尔X论V文Y网wwW.yOueRw.com 原文+QQ75201`8766

The Cre - LoxP recombination enzyme system is widely used in the target genes in mice, compared with the traditional gm technology, application of the system of gene knockout specificity of time and space, optimize the transgenic mouse model。 Application of Cre - LoxP recombination enzyme system construct of genetically modified mice can be in local implementation knockout mice, largely avoid premature death in mice, provide feasibility for follow-up study。

This experiment through genetic type building Sox2CreER transgenic mice, after expanding reproduction。 Then use PCR technology appraisal out successful build Sox2CreER transgenic mice, for subsequent Cre - LoxP recombination experiment to provide material support

Key words: transgenic mice ; PCR amplification ; Genotype identification

目    录

摘    要 2

Abstract 3

目    录 4

一． 引 言 5

1。1 背景介绍 5

二． 实验试剂与仪器 6

2。1 实验仪器 6

2。2 实验试剂 7

三．实验方法 8

3。1 DNA采集与提取 8