AUTOMATED SYNTHESIS OF LARGE DNA FRAGMENTS

大 DNA 片段的自动合成

• 批准号: 6211937
• 负责人: John Thomas Mulligan
• 金额: $ 12.71万
• 依托单位: BLUE HERON BIOTECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6211937
• 关键词: biomedical automation; biomedical equipment development; computer program /software; genetic manipulation; molecular cloning; nucleic acid chemical synthesis; oligonucleotides; technology /technique development

Current methods for the synthetic manufacture of large fragments of DNA, a process commonly referred to as "gene synthesis," are expensive, labor- intensive and slow. We propose to develop a novel automated, solid-phase system for gene synthesis that will reduce costs and speed delivery times. Solid-phase synthesis has already been shown to work on a limited scale for constructing DNA fragments of approximately 300 bp in a non-automated fashion. We plan to apply a team of scientists with expertise in molecular biology, nucleic acid chemistry, software development, polymer chemistry and automation to the problem of developing and optimizing solid-phase gene synthesis. Our goal is a robust automation system that will rapidly and inexpensively produce gene-sized fragments of DNA (up to 10,000 bp in length). In Phase I we will focus on developing prototype procedures for solid-phase gene synthesis that are amenable to automation. We will also develop the computer software needed to design and order the oligonucleotides that will be used to assemble the desired fragment of DNA. In Phase II we will develop the instrumentation and software needed for the automation of gene synthesis. PROPOSED COMMERCIAL APPLICATIONS: The commercial applications for cost-effective synthesis of large DNA fragments are extensive. A major bottleneck in many biological research programs is the time-consuming process of cloning and modifying genes to prepare them for detailed analysis. Rapid access to custom-engineered genes could replace a substantial fraction of the total cloning effort in industrial research groups and academic laboratories.

目前用于合成大片段DNA的方法（通常称为“基因合成”的过程）昂贵、劳动密集且缓慢。我们建议开发一种新型自动化固相基因合成系统，以降低成本并加快交付时间。固相合成已被证明可以在有限的规模上以非自动化方式构建约 300 bp 的 DNA 片段。我们计划运用一支在分子生物学、核酸化学、软件开发、高分子化学和自动化方面具有专业知识的科学家团队来解决固相基因合成的开发和优化问题。我们的目标是建立一个强大的自动化系统，能够快速、廉价地生产基因大小的 DNA 片段（长度最多 10,000 bp）。在第一阶段，我们将重点开发适合自动化的固相基因合成原型程序。我们还将开发设计和订购寡核苷酸所需的计算机软件，这些寡核苷酸将用于组装所需的 DNA 片段。在第二阶段，我们将开发基因合成自动化所需的仪器和软件。拟议的商业应用：经济高效地合成大 DNA 片段的商业应用非常广泛。许多生物研究项目的一个主要瓶颈是克隆和修改基因以准备进行详细分析的耗时过程。快速获得定制工程基因可以取代工业研究小组和学术实验室中克隆工作的很大一部分。