GENE MAPPING AND ALLELIC ASSOCIATIONS WITH CANCER

长期目标是建立一个基本的连锁图 实验室有袋类动物家养单胞虫的基因组，因此 为了增加这种动物模型的科学价值， 基础研究和应用研究情况。该基因的建立 MAP(任何有袋类动物的第一个MAP)将使基因定位成为可能 导致复合体在生理上相关的变异 发育异常和疾病等特征 易感性，并将提供关于 哺乳动物基因组的进化，特别是基因的保守性 Monodelphis与人类之间的共鸣和连锁组织。这 知识将有助于染色体定位和随后的 人类基因的鉴定和分子特征 基因组可能是基因的同源物，这些基因被鉴定为 Monodelphis模型中的疾病相关因素。 我们近期的计划是利用基因图谱来确定 候选抑癌基因和癌基因座的等位基因与 紫外线照射致皮肤和眼部肿瘤的发生 (黑色素细胞性痣、黑色素瘤和角膜肉瘤)，将进行研究 在本计划项目的项目1、2和3中。在未来，我们打算 将地图与项目4和项目4中生成的信息结合使用 5关于胆固醇代谢变异的遗传， 定位和识别影响差异脂血症的基因 对高脂肪、高胆固醇饮食挑战的反应。 在当前的项目期内，我们的目标是：1)生成一个 至少由100个匿名多态目录号码标记组成的地图 Monodelphis基因组中的序列(使用RAPD-PCR技术)，2)到 鉴定(通过Southern杂交/RFLP分析) 人类肿瘤抑制基因(APC、DCC、MCC、NF、 P53、Rb、WT)和ras家族癌基因(H-ras、K-ras、N-ras)，3)至 建立与随机扩增、肿瘤相关的连锁基因簇的同源性 抑制基因、癌基因和(先前已鉴定的)同工酶基因座 多重杂合子雌性单峰鱼的重组分析 表现出低重组率)，4)启动基因序列研究 和利用重组的合成标记基因座之间的连锁距离 来自多个杂合子男性(表现出高重组率)的数据 比率)，5)确定肿瘤表型和 抑癌基因和癌基因座上的特殊等位基因，6)到 确定所选肿瘤是否发生杂合性丢失(LOH) 黑色素瘤和角膜来源的培养细胞中的抑制基因 这一物种的肉瘤。

The long-term objectives are to establish a basic linkage map of the genome of the laboratory marsupial, Monodelphis domestica, and, thereby to increase the scientific value of this animal model in a variety of basic and applied research situations. The establishment of this gene map (the first for any marsupial) would enable the localization of genes that contribute to physiologically relevant variation in complex characteristics such as developmental anomalies and disease susceptibilities, and would provide additional insights regarding the evolution of the mammalian genome, particularly the conservation of gene syntony and linkage organization between Monodelphis and humans. This knowledge would facilitate the chromosomal localization and subsequent identification and molecular characterization of genes in the human genome that are possible homologs of genes that become identified as disease-related factors in the Monodelphis model. Our immediate plans are to utilize the gene map to identify associations between alleles at candidate tumor suppressor and oncogene loci and the occurrence of ultraviolet radiation-induced skin and eye neoplasia (melanocytic nevus, melanoma, and corneal sarcoma) that will be studied in Projects 1,2, and 3 of this Program Project. In the future we intend to use the map in concert with information generated in Projects 4 and 5 concerning the inheritance of variation in cholesterol metabolism, to localize and identify genes that affect differential lipemic responsiveness to high fat, high cholesterol dietary challenge. For the current project period, our aims are: 1) to generate a "framework map" composed of a minimum of 100 anonymous polymorphic DN marker sequences (using RAPD-PCR technology) in the Monodelphis genome, 2) to identify (via Southern blot/RFLP analysis) genetic polymorphisms among marsupial homologs of human tumor suppressor genes (APC, DCC, MCC, NF, p53, RB, WT) and ras family oncogenes (H-ras, K-ras, N-ras), 3) to establish the identities of linked gene clusters involving RAPD, tumor suppressor, oncogene, and (previously identified) isozyme loci via recombination analysis in multiply heterozygous female Monodelphis (which exhibit low recombination rates), 4) to initiate studies of gene order and linkage distances among synthetic marker loci using recombination data from multiply heterozygous males (which exhibit high recombination rates), 5) to identify associations between neoplastic phenotypes and particular alleles at tumor suppressor gene and oncogene loci, 6) to determine whether loss of heterozygosity (LOH) occurs at selected tumor suppressor loci in cultured cells derived from melanomas and corneal sarcomas of this species.