【作者】 乔丽平；

【导师】 杨德吉；

【作者基本信息】 南京农业大学 ， 临床兽医学， 2007， 硕士

【摘要】 微卫星不稳定性(microsatellite instability，MSI)广泛存在于人类肿瘤的基因组中。鸡马立克氏病(MD)是鸡的一种危害最严重的病毒所致的肿瘤性疾病，目前国内外鲜有关于鸡马立克肿瘤发生后宿主基因组的微卫星不稳定性的报道，其他家畜肿瘤上也没有宿主基因组MSI的研究报告。通过检测鸡胚成纤维细胞感染马立克氏病超强毒RBlB株后，鸡三号染色体上的基因组微卫星不稳定性情况，旨在发现是否存在与鸡马立克氏病发生相关的宿主基因组改变，为从宿主的角度研究鸡马立克肿瘤发生和发展的生物学提供基础研究资料。在本实验中应用27个SPF鸡胚，培育鸡胚成纤维细胞，来自同一鸡胚个体的成纤维细胞一半接种马立克氏病超强毒RBIB株，另一半不接毒，作为对照。分别提取培养细胞的基因组DNA，选取鸡三号染色体上27个微卫星标记，分别采用聚合酶链式反应、PAGE电泳、银染等技术检测基因组微卫星不稳定性。结果，27个微卫星位点中有20个出现微卫星不稳定性，平均变化频率为工6。30％，变化频率最高的为ABRl41、MCW259、ABR426(25.93％)；没有检测到杂合性缺失。24例鸡胚有两个或两个以上位点出现基因组变化。本研究首次发现鸡胚成纤维细胞感染马立克氏病病毒后3号染色体上存在多位点的微卫星不稳定性。感染马立克氏病病毒后CEF可出现多个微卫星位点的改变，提示MDV感染CEF早期可能会整合到宿主的多个位点引起基因组的变化。以PCR、电泳和银染为基础的微卫星技术可以快速、有效地检测出染色体内部基因组的不稳定性，这为动物肿瘤疾病的研究提供了一个很好的癌基因研究模式。更多还原

【Abstract】 Microsatellite instability(MSI) has been proved to exist widely in the genome oftumor of human. MD, one of the chicken tumor disease caused by MDV, harmoverwhelmingly. At home and abroad, the MSI of host genome are reported rarelywhen MD tumor emerge, and neither are for other domestic animal tumor. Thatinvestigate microsatellite instability of Chromosome 3 of the chicken embryofibroblasts infected with the virulent MDV RB1B strain aims to provide basic data forbiology of formation and development of Marek’s disease in termes of host genome.In this study,chicken embryo fibroblasts were prepared from 27 SPF chickenembryos. The half of CEF derived from each SPF chicken embryo were infected withthe virulent MDV-RB1B strain, the others were used as control. Their genomic DNAwas extracted, respectively. MSI of Chromosome 3 were detected at 27microsaterllite loci by means of PCR,PAGE electrophoresis and silver staining inmatched normal CEF and MDV-infected CEE MSI were found on 20 of 27 loci andthe average MSI frequency was 16.93%. MSI loci with the highest frequencies of29.63% were identified at ABR141、MCW259 and ABR426; LOH was not found.MSI were found in 24 cases at two or more than two loci.In this study, it is first foundmicrosatellite instabilities on Chromosome 3 of CEF infected with MDV-RB1B strain.Finding MSI at several microsatellite loci possibly means that the genes of MDVintegrate into the genome of the host and cause the genome changes of the host in theearlier period of the MDV infecting CEF.MSI detection,based on PCR、electrophoresis and silver staining can quickly andefficiently discover the MSI of the genome. It provides a mode for the research on thecancer gene of other animal tumor disease.更多还原