Xiphophorus Gene Map Saturation--Genes Regulating Tumor Development

剑尾鱼基因图谱饱和--调控肿瘤发展的基因

• 批准号: 6259620
• 负责人: DONALD C MORIZOT
• 金额: $ 27.06万
• 依托单位: TEXAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2004-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6259620
• 关键词: Osteichthyes; alternatives to animals in research; animal genetic material tag; artificial chromosomes; carcinogenesis; genetic markers; genetic models; linkage mapping; model design /development; neoplasm /cancer genetics; oncogenes; tumor suppressor genes

DESCRIPTION: (Applicant's Description) The gene map of Xiphophorus fishes (Teleostei: Poeciliiidae) is one of the largest among vertebrates, with more than 100 genes among 350 markers assigned to independently assorting multipoint linkage groups residing on probably all 24 acrocentric chromosome pairs. This detailed linkage map has been used to identify a specific sex chromosome-linked oncogene and an autosomal CDNK2 tumor suppressor gene associated with susceptibility to spontaneous or UV-induced melanoma formation in interspecific backcross hybrids. Goals of this proposal include: 1) expansion of the gene map to more than 200 loci by mapping cellular oncogenes, potential tumor suppressor genes, cell cycle control genes, and members of other multigene families; 2) to saturate the linkage map with more than 500 anonymous DNA markers, primarily utilizing arbitrarily primed polymerase chain reaction (AP-PCR) techniques to generate polymorphic amplified DNA sequences; and 3) to increase the number of sequence tagged products in the Xiphophorus genome to facilitate positional cloning of genes of interest and to relate recombination distance to physical distance in the genome. A number of genetic crosses, each consisting of more than 100 backcross hybrids, will be utilized to assess linkage relationships of all informative genetic markers, including isozyme, pigment pattern, DNA restriction fragment length (RFL), specifically primed PCR amplification, and AP- PCR polymorphisms. Initially, most crosses will segregate approximately 100 markers which will establish most of the multipoint linkage groups. Linkage of markers in multiple crosses coupled with expansion of numbers of informative polymorphisms will finally coalesce multipoint linkage groups into 24 chromosomal linkage groups. By the end of the project period, placement of approximately 750 markers will result in an average of a marker every 2-5 centimorgans, and will provide many opportunities to correlate physical distances of markers in one or contiguous YAC segments with recombination frequencies. The most important contribution of a densely marked genetic map will be the potential to detect through linkage analyses new tumor regulatory genes in Xiphophorus models. Addition of a large number of characterized genes may provide candidates for such regulatory, or alternatively, positional cloning may identify previously uncharacterized regulatory genes whose homologues control tumorigenesis in man.

产品说明：剑尾鱼（真骨鱼纲：Poeciliidae）的基因图谱是脊椎动物中最大的基因图谱之一，在350个标记中有100多个基因被分配给独立的多点连锁群，这些多点连锁群可能位于所有24个近端着丝粒染色体对上。这个详细的连锁图已被用来确定一个特定的性染色体连锁癌基因和常染色体CDNK 2肿瘤抑制基因与易感性自发或紫外线诱导的黑色素瘤形成种间回交杂交。该计划的目标包括：1）通过定位细胞癌基因、潜在的肿瘤抑制基因、细胞周期控制基因和其他多基因家族的成员，将基因图谱扩展到200多个位点; 2）用500多个匿名DNA标记使连锁图谱饱和，主要利用任意引物聚合酶链反应（AP-PCR）技术产生多态性扩增DNA序列;和3）增加剑尾鱼基因组中序列标记产物的数量，以促进感兴趣基因的定位克隆，并将重组距离与基因组中的物理距离相关联。将利用多个遗传杂交，每个杂交由100个以上的回交杂种组成，以评估所有信息性遗传标记的连锁关系，包括同工酶、色素模式、DNA限制性片段长度（RFL）、特异性引物PCR扩增和AP-PCR多态性。最初，大多数杂交将分离大约100个标记，这些标记将建立大多数多点连锁群。多个杂交中标记的连锁加上信息多态性数目的扩大，最终将多点连锁群合并为24个染色体连锁群。到项目期结束时，放置大约750个标记将导致平均每2-5厘摩一个标记，并将提供许多机会将一个或连续YAC片段中标记的物理距离与重组频率相关联。密集标记的遗传图谱的最重要的贡献将是通过连锁分析在剑尾鱼模型中检测新的肿瘤调控基因的潜力。添加大量的特征基因可以提供这样的调节的候选者，或者，定位克隆可以鉴定以前未表征的调节基因，其同源物控制人类肿瘤发生。