ISOLATION OF A HUMAN HOMOLOGUE OF THE MURINE LY-6 SYSTEM

鼠Ly-6基因座编码一组细胞表面分子 在多种淋巴样、髓样和其它细胞类型上表达。 这些Ly-6编码分子中的至少两种，Ly-6E.1和TAP， 并参与T淋巴细胞活化。 一个cDNA的一个 Ly-6蛋白是近年来分离到的一种蛋白质， 证实Ly-6是一个复杂的多基因家族。 我们建议 使用鼠cDNA分离和表征人 小鼠Ly-6系统的同源物。 人的存在 与Ly-6同源的序列将通过探测 人基因组DNA与鼠Ly-6的Southern印迹 cDNA。 表达与Ly-6同源的mRNA的细胞将是 通过用Ly-6 cDNA探测北方印迹来鉴定。 cDNA 来自表达Ly-6相关信息的细胞的文库将被 用Ly-6探针筛选同源克隆， 与世隔绝 这些人cDNA的特征在于： 限制性酶切分析并测序。 肽将 基于推导的氨基酸序列合成， 用于产生特异性抗体。 全长cDNA将 在表达载体中表达，并且分离的蛋白质将 用于结构研究和生成特定的 抗体的 这些cDNA和抗体将用于 研究人类Ly-6系统的结构及其作用， 在细胞激活中起作用。 这项工作的主要长期目标是确定和 表征参与转导过程的分子 细胞外信息和细胞激活。 它已成为 最近清楚，粘附功能或细胞的控制 活化分子可能是某些或许多人类疾病的病因 恶性肿瘤。 此外，活化的淋巴细胞是一种重要的 治疗某些人类恶性肿瘤的新模式。 阐明人类Ly-6系统将进一步我们的研究。 了解癌症并帮助其治疗。

The murine Ly-6 locus encodes a group of cell surface molecules expressed on a variety of lymphoid, myeloid and other cell types. At least two of these Ly-6 encoded molecules, Ly-6E.1 and TAP, and involved in T lymphocytes activation. A cDNA for one of the Ly-6 proteins was isolated recently and studies with this cDNA confirm that Ly-6 is a complex multigene family. We propose to use the murine cDNA to isolate and characterize a human homologue of the murine Ly-6 system. The existence of human sequences homologous to Ly-6 will be determined by probing Southern blots of human genomic DNA with the murine Ly-6 cDNA. Cells expressing mRNA homologous to Ly-6 will be identified by probing Northern blots with the Ly-6 cDNA. cDNA libraries from cells expressing Ly-6 related messages will be screened with the Ly-6 probe and homologous clones will be isolated. These human cDNA's will be characterized by restriction analysis and will be sequenced. Peptides will be synthesized based on the deduced amino acid sequences and will be used to generate specific antibodies. Full length cDNA's will be expressed in expression vectors and the isolated proteins will be used for structural studies and for the generation of specific antibodies. These cDNA's and antibodies will be utilized for studying the structure of the human Ly-6 system and the role it plays in cellular activation. The major long-term objective of this work is to identify and characterize molecules involved in the process of transducing extracellular messages and cellular activation. It has become clear recently that adherent function or control of cellular activation molecules may be an etiology for some or many human malignancies. Moreover, activated lymphocytes are an important new modality in the treatment of some human malignancies. Elucidating the human Ly-6 system should further our understanding of cancer and aid in its treatment.