MOLECULAR BIOLOGY OF LYMPHOCYTE AND NEURONAL GROWTH

描述：该计划的总体目标是获得一个完整的 尽可能多地了解在 B淋巴细胞的分化，并开始将分析扩展到 神经系统。这项工作对研究 自身免疫性疾病、免疫反应、免疫记忆和神经元 赤字。下一个授权期的具体目标是理解 单个转录增强子的作用，内含子增强子 Kappa(K)轻链；了解免疫球蛋白基因重排 生物化学水平上的过程及其控制；开发一种标记的方法 记忆细胞和扩展免疫分化的分析方法 对神经元分化的研究。Kappa基因表达被认为是 由2个增强剂控制。这些增强剂在体内的各种作用 将使用同源重组来分析免疫分化 在ES细胞中制造关键突变，然后可以用来重建 RAG-2基因缺失小鼠的淋巴系统。ES细胞同源重组 还将用于在L基因中插入K调控元件以 检查小鼠K/L轻链比率是否为 两者转录调控元件的相对优势 轻链基因。免疫球蛋白基因重排过程是一个 独特的DNA重排过程。为了研究它，各种相关的 蛋白质将被提纯，并将检测它们的体外活性。 这一分析将扩展到染色质和甲基化DNA，以尝试 了解重排过程的不同级别的控制。 对细胞的分析将被用来揭示更复杂的控制水平。 记忆B细胞和T细胞表面没有明确的标记。 将产生转基因小鼠，其中记忆B和T细胞将被 基因标记的，因此可以明确地识别和检查。 免疫分化的研究得益于可获得的 克隆细胞系和嵌合分析方法。要扩展这些功能 有利于神经元分化的研究，方法将会发展 利用癌基因制备分化神经元的克隆性衍生物 精确的贴标签程序。也是为了研究在其他方面致命的作用 基因在神经元发育中，嵌合分析的方法将是 发展起来的。它将特别应用于分析 神经系统信号转导中的核因子-KB转录因子。

DESCRIPTION: The overall aim of this program is to gain as complete a knowledge as possible of the events that transpire during the differentiation of B lymphocytes and to begin to extend the analysis to the nervous system. This work is particularly relevant to the study of autoimmune disease, immunologic responses, immunologic memory and neuronal deficits. The specific aims of the next granting period are to understand the role of a single transcriptional enhancer, the intron enhancer of the Kappa (K) light chain; to understand the immunoglobulin gene rearrangement process and its control at a biochemical level; to develop a method to mark memory cells and to extend the methods of analysis of immunodifferentiation to the study of neuronal differentiation. Kappa gene expression is thought to be controlled by 2 enhancers. The various roles of these enhancers in immunodifferentiation will be analyzed using homologous recombination to make critical mutants in ES cells that can then be used to reconstitute the lymphoid systems of RAG-2-deleted mice. ES cell homologous recombination will also be used to insert K regulatory elements into the l genes to examine whether the K/l light chain ratio in the mouse is a consequence of the relative strengths of the transcriptional regulatory elements of the two light chain genes. The immunoglobulin gene rearrangement process is a unique process of DNA rearrangement. To study it, the various relevant proteins will be purified and their in vitro activities will be examined. This analysis will be extended to chromatin and methylated DNA to attempt to understand the various levels of control of the rearrangement process. Analysis in cells will be used to uncover more complex levels of control. There is no unambiguous marker on the surface of memory B and T cells. Transgenic mice will be generated in which the memory B and T cells will be genetically marked and thus can be identified and examined unambiguously. The study of immunodifferentiation has benefitted from the availability of clonal cell lines and methods of chimeric analysis. To extend these benefits to the study of neuronal differentiation, methods will be developed to prepare clonal derivatives of differentiated neurons using oncogenes and a precise labeling procedure. Also to study the role of otherwise lethal genes in neuronal development, a method of chimeric analysis will be developed. It will be applied particularly to analyzing the role of the NF-KB transcription factor in nervous system signaling.