Establishment of Detection Systems and Expression Systems for Organ-Specific Mucins

点击翻译按钮获取中文摘要

The aims of this proposal is to establish systems to detect various human epithelial mucins and to establish systems to obtain stable transfectant cells expressing mucin core polypeptide cDNA.Specifically, preparation of monoclonal antibodies specific for unique mucins and preparation of expression vectors containing mucin core polypeptide cDNA were conducted. Elucidation of the transcriptional regulation of mucin core polypeptides was also performed.The following list consists of the findings obtained during the project term : (1) A novel monoclonal antibody MY.1E12 specific for sialylated MUC1 mucin was established. (2) Vectors containing chimera mucins (MUC1/MMGL and MUC2/MMGL) have been prepared. (3) A stable transfectant cell line and clones have been prepared using human renal carcinoma cells. The biological behavior of these cells were investigated. (4) Glycosylation of MUC1 mucins expressed by transfection of K562 human erythroleukemia cells was compared to the glycosylation of endogenous leukocyte mucins. (5) A lectin expressed by a strain of enterobacterium was isolated and shown to bind MUC2 mucin but not MUC1 mucin. (6) A novel lecting that was isolated from this bacterium. (7) An inducible transcriptional cis-acting element for MUC1 mucin gene has been identified. Binding proteins to this element were isolated from nuclear extracts of mammary carcinoma cells. (8) A cDNA corresponding to human macrophage lectin has been cloned and expressed. The carbohydrate binding specificity of this lectin has been demonstrated to bind to Tn and sialyl-Tn antigens.