AUTOMATED SYNTHESIS OF LARGE DNA FRAGMENTS

目前合成大片段DNA的方法，通常被称为“基因合成”，都是昂贵的、劳动密集型的，而且速度很慢。我们建议开发一种新的自动化、固相基因合成系统，以降低成本并加快交付时间。固相合成已经被证明在有限的规模上工作，以非自动化的方式构建大约300bp的DNA片段。我们计划应用一支在分子生物学、核酸化学、软件开发、聚合物化学和自动化方面具有专业知识的科学家团队来开发和优化固相基因合成。我们的目标是一个强大的自动化系统，它将快速而廉价地产生基因大小的DNA片段(长度可达10,000个碱基对)。在第一阶段，我们将专注于开发适合自动化的固相基因合成的原型程序。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：经济有效地合成大DNA片段的商业应用非常广泛。许多生物学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能会取代工业研究小组和学术实验室总克隆工作的很大一部分。

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.