MONOCLONAL ANTIBODY PROBES FOR A, B, AND Z HELICES

A、B 和 Z 螺旋的单克隆抗体探针

• 批准号: 3292132
• 负责人: DAVID A ZARLING
• 金额: $ 3.79万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 1986-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3292132
• 关键词: acetylaminofluorene; antibody formation; antibody specificity; chemical carcinogen; environmental stressor; gel electrophoresis; gel filtration chromatography; immunologic techniques; nucleic acid sequence; radiotracer; scintillation spectrometry; tissue /cell culture

The goal of the proposed research is to produce and characterize monoclonal antibodies against antigenic determinants on biologically relevant double-stranded DNA and RNa conformations and sequences. The primary research objective is to use mouse monoclonal antibody probes to distinguish unique as well as common structural determinants on A, B, and Z double-stranded nucleic acid helices. These structures include antigenic features of the grooves, phosphate backbones, or bases in A, B, or Z helices. This immunochemical analysis of DNA and RNA helices is of fundamental importance in understanding features of double-helices present in physiological, environmentally stressed, or chemical carcinogen modified sequences recognized by enzymes or proteins. The specific aims are: (1) to prepare and characterize new monoclonal antibodies against antigens of the synthetic linear Z-poly[d(A-C) d(G-T)] family which we have recently stabilized in a Z state under physiological ionic conditions, (2) to continue studying common and unique antigenic determinants on left-handed Z DNA and Z RNA sequences using monoclonal antibodies prepared against Z RNA (e.g. br-poly[r(G-C)] immunogens), (3) to continue the immunological analysis of B DNA and A RNA sequences using enzymatically and/or chemically synthesized sequences as immunogens for in vitro and in vivo immunizations and as substrates for immunoassay, (4) to determine the effects of cations, drugs, and carcinogens (e.g. N-acetoxy-2-acetylaminofluorene) on antibody recognized Z conformations and (5) as a longer-term aim to introduce these monoclonal antibodies to A, B, and Z helices into living cells (e.g. by microinjection) and determine their various distributions and potential functions.

这项研究的目的是生产和表征单克隆抗体， 生物学相关抗原决定簇抗体 双链DNA和RNa构象和序列。 主 研究目的是利用小鼠单克隆抗体探针， 区分A、B和Z上的独特和共同结构决定因素 双链核酸螺旋。 这些结构包括抗原 A、B或Z中的凹槽、磷酸骨架或碱基的特征 螺旋 这种DNA和RNA螺旋的免疫化学分析， 在理解双螺旋存在的特征方面的根本重要性 在生理、环境应激或化学致癌物修饰中， 由酶或蛋白质识别的序列。 具体目标是：（1） 制备和表征针对以下抗原的新的单克隆抗体： 合成线性Z-聚[d（A-C）d（G-T）]家族， 在生理离子条件下稳定在Z状态，（2）至 继续研究左手Z的共同和独特抗原决定簇 使用针对Z RNA制备的单克隆抗体进行DNA和Z RNA序列测定 (e.g. br-聚[r（G-C）]免疫原），（3）继续免疫 使用酶和/或化学方法分析B DNA和A RNA序列 作为体外和体内免疫的免疫原的合成序列 和作为免疫测定的底物，（4）测定阳离子的作用， 药物和致癌物（如N-乙酰氧基-2-乙酰氨基芴） 认识到Z构象和（5）作为一个长期的目标，介绍这些 将针对A、B和Z螺旋的单克隆抗体插入活细胞（例如通过 显微注射），并确定它们的各种分布和潜力 功能协调发展的