微卫星DNA在近交系豫医无毛小鼠遗传背景监测中的应用

发布时间：2018-03-21 09:49

本文选题：聚合酶链反应　切入点：豫医无毛小鼠　出处：《郑州大学》2006年硕士论文　论文类型：学位论文

【摘要】：目的：采用高度近交的动物可以减少实验动物数量和实验重复次数，提高实验结果的准确性、可比性和可重复性。近交系动物的显著特征是基因纯合性及遗传稳定性，但在培育、保种及繁殖生产过程中，存在着发生遗传变异或遗传污染的可能，所以，需要对近交系小鼠进行严格、长期、定时的遗传监测。传统的皮肤移植、生化标记等通过检测其表型变化来推测其基因变化的方法有明显的局限性，而微卫星DNA多态性高、分布广、数目多、比较稳定、易于扩增，可直接检测DNA水平基因变化，能够更全面地反映基因组的遗传概貌及变异情况，且微卫星DNA具有检测灵敏、特异、简便、省时等优点，所以，它将成为近交系动物较理想的遗传监测标记。豫医无毛小鼠(Yuyi Hairless Mice，简称YYHL小鼠)是河南省实验动物中心1990年在昆明小鼠繁育过程中发现的小鼠突变种群，是不同于裸鼠的一种皮肤和毛发结构发生遗传突变的新品系。近交系YYHL小鼠被广泛应用于组织学、肿瘤病因学和肿瘤治疗等方面，但在皮肤老化、脱发等皮肤病变及化妆品、外用药品的检测等临床研究中更具有代表性，是非常理想的实验材料。但有关近交系YYHL小鼠在分子生物学遗传监测方面的研究较少。本实验采用18对引物对近交系小鼠的DNA多态性进行研究：提取小鼠基因组DNA并利用微卫星引物PCR扩增，通过琼脂糖凝胶电泳、图像分析仪观察有无扩增产物，然后再通过聚丙烯酰胺凝胶电泳及银染等技术显示微卫星DNA扩增产物。最后采用遗传学数据分析方法进行分析，以期筛选出具有较高多态性的微卫星位点，用于近交系小鼠的遗传背景监测。方法：选用七品系近交系小鼠(BALB／C、YYHL小鼠、C57、CBA、DBA／2、C3H／HeJ、
[Abstract]:Objective: using highly inbred animals can reduce the number of experimental animals and increase the accuracy, comparability and repeatability of experimental results. The significant characteristics of inbred animals are genetic homozygosity and genetic stability. However, in the process of breeding, preserving and breeding, there is the possibility of genetic variation or genetic contamination. Therefore, it is necessary to carry out strict, long-term and regular genetic monitoring of inbred mice. Biochemical markers and other methods to detect phenotypic changes have obvious limitations, while microsatellite DNA is highly polymorphic, widely distributed, numerous, stable and easy to amplify, and can be directly detected for gene changes at DNA level. Microsatellite DNA has the advantages of sensitivity, specificity, simplicity and time saving. It will be an ideal genetic monitoring marker for inbred animals. Yuyi Hairless Mice (YYHL mouse) is a mutated mouse population discovered in Kunming mouse breeding process in 1990 by Henan Provincial Laboratory Animal Center. Inbred YYHL mice are widely used in histology, tumor etiology and tumor therapy, but in skin aging, alopecia and other skin diseases and cosmetics. The detection of external medicine is more representative in clinical research, and it is a very ideal experimental material. In this study, 18 pairs of primers were used to study the DNA polymorphism of inbred YYHL mice. Genomic DNA was extracted from mice and amplified by microsatellite primer PCR. Agarose gel electrophoresis and image analyzer were used to observe the amplification products, and then polyacrylamide gel electrophoresis and silver staining were used to show the microsatellite DNA amplification products. The aim of this study was to screen microsatellite loci with high polymorphism for genetic background monitoring in inbred mice. Methods: seven inbred strain mice, BALB / Cnyyhl mice, C57 / CBA / DBA / 2 / C3H / HeJ, were used.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R-332

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