BUDR DEPENDENCE, MALIGNANCY, AND DIFFERENTIATION

这项研究的长期目标是阐明分子 黑色素瘤细胞分化的调控机制。这个 实验将涉及体细胞遗传学技术(CELL 杂交和基因转移)和胸腺嘧啶核苷类似物5- 溴脱氧尿苷(BrdU)与 重组DNA。 这项研究的具体目的与灭绝有关 体细胞杂交中的色素沉着，成纤维细胞的转运 杂交种中的酪氨酸酶基因，以及BrdU对色素沉着的抑制。 细胞杂交的实验将集中在染色体上。 导致物种灭绝的调控因素的本地化， 确定灭火器的位置，发生 反式激活，识别DNA序列(假设 上游)参与调控的酪氨酸酶基因，以及 对这些序列影响基因表达方式的分析。这个 BrdU的实验将集中在DNA序列的鉴定上 在参与抑制的酪氨酸酶基因中，分析 这些序列的作用方式，以及角色的确定 BrdU在调节因子与调节因子相互作用中的作用 酪氨酸酶基因的区域。这些实验应该会导致 确定涉及顺式和反式作用的调控元件 酪氨酸酶基因表达的调控对细菌鉴定的影响 结合这些元素或由这些元素产生的因子，以及与 灭火器的轨迹。 这些实验应该有助于阐明分子。 哺乳动物细胞分化的调控机制。这个 实验也将有助于我们对 黑色素瘤细胞。所获得的关于监管机制的信息可能 与肿瘤发生有关，因为恶性肿瘤(如分化)是 受细胞杂交和BrdU的影响。

The long term goal of this research is to elucidate the molecular mechanisms by which differentiation is regulated in melanoma cells. The experiments will involve the techniques of somatic cell genetics (cell hybridization and gene transfer) and the use of the thymidine analog 5- bromodeoxyuridine (BrdU) in combination with the techniques of recombinant DNA. The specific aims of the research relate to the extinction of pigmentation in somatic cell hybrids, the transaction of the fibroblast tyrosinase gene in hybrids, and the suppression of pigmentation by BrdU. The experiments with cell hybrids will focus on the chromosomal localization of the regulatory element causing extinction, the identification of the extinguisher locus, the occurrence of transactivation, the identification of DNA sequences (presumably upstream) in the tyrosinase gene that are involved in regulation, and the analysis of the way in which these sequences affect gene expression. The experiments with BrdU will focus on the identification of DNA sequences in the tyrosinase gene that are involved in suppression, the analysis of the way in which these sequences act, and the determination of the role of BrdU in the interactions between regulatory factors and regulatory regions of the tyrosinase gene. These experiments should lead to the identification of cis- and trans-acting regulatory elements involved in the regulation of tyrosinase gene expression, to the identification of factors binding to or produced by these elements, and to the isolation of the extinguisher locus. These experiments should contribute to the elucidation of the molecular mechanisms regulating differentiation in mammalian cells. The experiments also will contribute to our knowledge of the biology of melanoma cells. The information gained about regulatory mechanisms could be relevant to tumorigenesis, since malignancy (like differentiation) is affected by cell hybridization and by BrdU.