FILAMENTOUS FUSION PHAGE

丝状融合噬菌体

• 批准号: 3299719
• 负责人: GEORGE PEARSON SMITH
• 金额: $ 3.57万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3299719
• 关键词: DNA directed RNA polymerase; Salmonella; antibody specificity; antigens; bacterial virus; epitope mapping; genetic library; genetic manipulation; hemerythrin; molecular cloning; synthetic peptide

Filamentous fusion phage display the amino acids coded by a cloned DNA insertion the surface of an infectious virus particle. Phage bearing a particular foreign determinant can be affinity-purified with antibody, making it easy to purify clones that are as rare as 1 in 100 million in the original mixture. Methods will be devised for constructing fusion-phage "libraries" containing 100 million different foreign DNa inserts, so that the full potential of the technology can be exploited. The concept of an epitope library exemplifies the uses to which fusion phage might be put. The foreign DNA inserts in this library would be synthetic DNa with random sequence. Billions of short amino acid sequences would be represented in a 100-million-clone library; it is likely, therefore, that it will contain short determinants ("epitopes") recognized by any anti-protein antibody. This idea will be tested with antibodies directed against myohemerythrin, a small protein whose antigenic structure has been intensively studied with synthetic peptides. The results should yield a wealth of new information about the specificity of anti-protein antibodies-- information that touches on the fundamental issue of how the immune system manages to specifically recognize a seemingly unlimited repertoire of different protein antigens. They should also indicate the feasibility of future applications of the epitope library. One possibility is that it could be used to determine the epitope recognized by an interesting antibody--one, say, that confers protective immunity to some disease. This information could then be used to design a synthetic vaccine or immunogen capable of eliciting similar antibodies in other individuals--all without having to clone and analyze the gene encoding the natural antigen.

丝状融合噬菌体展示克隆编码的氨基酸 DNA 插入传染性病毒颗粒的表面。 噬菌体 带有特定外来决定簇的可以进行亲和纯化 带有抗体，可以轻松纯化罕见的克隆 原始混合物中的一亿分之一。 将制定方法 用于构建包含 1 亿个的融合噬菌体“文库” 不同的外源 DNA 插入片段，从而充分发挥 技术可以被利用。 表位库的概念 举例说明了融合噬菌体的用途。 这 该文库中的外源 DNA 插入片段将是合成 DNA 随机序列。 数十亿个短氨基酸序列将是 代表一亿个克隆库；很可能， 因此，它将包含短决定簇（“表位”） 被任何抗蛋白质抗体识别。 这个想法将被测试 含有针对肌红蛋白（一种小蛋白质）的抗体 其抗原结构已被深入研究 合成肽。 结果应该会产生大量新的 有关抗蛋白抗体特异性的信息—— 涉及免疫如何发挥作用这一基本问题的信息 系统设法专门识别看似无限的 不同蛋白质抗原的库。 他们还应该 表明该表位未来应用的可行性 图书馆。 一种可能性是它可以用来确定 表位被一种有趣的抗体识别——比如说， 赋予对某些疾病的保护性免疫力。 此信息 然后可用于设计合成疫苗或免疫原 能够在其他个体中引发类似的抗体——全部 无需克隆和分析编码天然物质的基因 抗原。