摘要小鼠作为哺乳动物遗传研究中首选的模式生物。经过过去几十年的努力，培育基因打靶和转基因小鼠已经变成很平常的工作，以目前的技术可以获得在核酸水平上的突变品系。现在，诸如此种基因组修饰变得越来越精细，而在基因水平上标记不同的细胞群已成为非常普遍的研究。52881

    随着人类基因组计划（HGP）的顺利完成，后基因时代的生物学研究迫切需要一种有效的基因功能分析方法。基因敲除小鼠模型的应用，为研究基因的功能和寻找新的治疗人类疾病的干预措施提供了有力支持。基因打靶和基因捕获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。

Lac Z是基因工程实验中的常用标记基因。GFP能够在体内或体外进行实时监测。在GFP中有两种全新颜色的变体ECFP和EYFP。由于ECFP和EYFP的谱线轮廓截然不同且没有重叠，所以这两种荧光蛋白可同时用作报告基因并共同显色，这对体内双标记或荧光能量转移分析等研究是非常理想的。GFP及其颜色变体作为报告蛋白的应用为科学研究提供了一个空前的高精度检测方法，代表着小鼠基因组改造技术的未来发展方向，并开启了在同一个动物体内应用组合无创性报告基因的广阔前景。

    因此本文将通过Isl1lacZ/Isl2GFP两种用于分析Isl1/Isl2基因表达模型的标签小鼠的基因型鉴定。

毕业论文关键词：转基因小鼠； PCR扩增； 基因敲除 

Abstract Mouse as the preferred model organism in the study of mammalian genetics. After decades of efforts, cultivate gene targeting and transgenic mice have become very common, to the current technology can be obtained at the nucleic acid level of the mutant lines. Now, such as the modification of the genome is becoming more and more precise, and it has become very common to mark different cell populations at the gene level.

Z Lac is a commonly used marker gene in genetic engineering experiments. GFP can be real-time monitoring in vivo or in vitro. In GFP there are two different colors of the variant ECFP and EYFP. Because the ECFP and EYFP spectral line profile distinct and there is no overlap, so the two fluorescent protein can be also used as report gene and common color, which the body double labeling or fluorescence energy transfer analysis and so on is very ideal. GFP and its color variant application as a reporter protein to the scientific research provides an unprecedented high precision detection method, represent the future development direction of the mouse genome transformation technology, and open the noninvasively the broad prospects of the reporter gene in the same animal using the combination.

Therefore, this paper will be used to analyze the identification of the gene expression model of Isl1/Isl2 gene in mice by Isl1lacZ/Isl2GFP.

Key words: transgenic mice； PCR amplification；Gene knockout

目录

摘  要 2

Abstract 3

目  录 4

第一章 引言 5

1.1 基因简介 5

第二章 实验材料与方法 6

2.1实验材料 6

2.1.1实验试剂 6

2.1.2实验仪器 6

2.2 实验方法 7

2.2.1 小鼠饲养 7

2.2.2 样品采集 9

2.2.3 小鼠DNA的提取 9

2.2.4  Thermo ND2000C测量DNA浓度