摘要目的 视觉功能是人体最重要的感觉功能之一,人从外界获取的信息有80%是通过眼睛来实现的。视力障碍从多方面影响人们的正常生活。在研究视网膜发育和疾病中,Sox2在早期胚胎的神经管表达,随后在视网膜和神经中枢表达,对早期胚胎发生、神经发生等重要的发育事件中起重要作用运用,但是我们对Sox2的了解还远远不够,需要我们进一步对Sox2进行研究。本实验运用PCR鉴定已经构建完成的Sox2-CreER转基因小鼠的基因型,为之后用解剖学和分子生物学技术检验Sox2-CreER转基因小鼠的发育状况,确定Sox2-CreER转基因小鼠是预期的基因型和正常的发育状况等进一步实验做铺垫工作。同时根据鉴定结果,采用Cre-loxP系统杂交Sox2-CreER转基因小鼠和R26RlacZ小鼠,跟踪Sxo2-CreER的时空表达和表达Sox2细胞的命运。方法 构建Sox2CreER转基因小鼠;DNA提取;聚合酶链式反应(PCR)扩增;凝胶电泳;结果 依据凝胶电泳图判断每只带有编号的小鼠是否转基因成功。47407
毕业论文关键词:Sox2;PCR扩增; 基因型鉴定;
Abstract:Object Visual function is one of the body's most important sensory function, personal information obtained from the outside world through the eyes of 80% to achieve. In the study of retinal development and disease, Sox2 expression in the early embryonic neural tube, followed by expression in the retina and nerve center of early embryogenesis, and play an important role in developmental events and neurogenesis and so on. But our understanding of Sox2 is not enough, We need to carry out further research on Sox2. This experiment has been constructed using PCR to identify the completed Sox2-CreER transgenic mice genotype. Is followed by anatomical and molecular biology test Sox2-CreER transgenic mice development status, determine Sox2-CreER transgenic mice is expected genotype and normal development status to pave the way for further experimental work. At the same time according to the identification results using Cre-loxP system hybrid Sox2-CreER transgenic mice and mice R26RlacZ track Sxo2-CreER spatial and temporal expression and the fate of Sox2 expression cells. Method Construction Sox2CreER transgenic mice; DNA extraction; polymerase chain reaction (PCR) amplification; gel electrophoresis; Result Sox2Cre gene has been successfully introduced into a variety of homozygous, heterozygous mice.
Keyword: Sox2;PCR increase;Genotyping
目录
1. 引言 5
1.1 视网膜组成及其信号传递 5
1.2 Sox2简介 6
1.2.1 Sox2 蛋白及cDNA 序列的特点 6
1.2.2 Sox2与早期胚胎发生 6
1.2.3 Sox2与神经发育 7
1.2.4 Sox2与晶状体发育 7
2. 实验内容与设计 7
2.1 实验目的与意义 7
2.2 实验内容 8
3. 试验试剂与方法 8
3.1 实验小鼠的饲养 8
3.2 样品采集 8