Reliable Assembly of Cloned DNA Fragments

克隆 DNA 片段的可靠组装

• 批准号: 7108928
• 负责人: John Thomas Mulligan
• 金额: $ 36.62万
• 依托单位: BLUE HERON BIOTECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-05 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7108928
• 关键词: DNA; DNA binding protein; DNA methylation; artificial chromosomes; binding sites; biotechnology; expression cloning; high throughput technology; macromolecule; method development; methyltransferase; molecular assembly /self assembly; nucleic acid chemical synthesis; plasmids; restriction endonucleases; transfection /expression vector

DESCRIPTION (provided by applicant): Our goal in this project is to develop a uniform method for assembling large DNA molecules by ligation of sticky-ended fragments, regardless of the sequence, the fragment length, or the presence of restriction enzyme sites. If we are successful, this technology could be used to assemble full-length cDNAs, mammalian genes, knockout constructs, gene clusters, and even whole BACs or bacterial genomes. The combination of a universal fragment assembly technology with Blue Heron's core technology, gene synthesis, will provide researchers with rapid, cost-effective access to any DNA sequence, regardless of size or sequence composition. It will speed the pace of research by allowing scientists to focus on experiments rather than cutting and pasting DNA molecules. It wll also enable new projects that would be impossible or impractically expensive without full control of large DNA sequences.

描述（由申请人提供）：我们在该项目中的目标是开发一种通过连接粘性末端片段来组装大 DNA 分子的统一方法，无论序列、片段长度或是否存在限制性酶位点。如果我们成功，这项技术可用于组装全长 cDNA、哺乳动物基因、敲除构建体、基因簇，甚至整个 BAC 或细菌基因组。通用片段组装技术与 Blue Heron 的核心技术基因合成相结合，将为研究人员提供快速、经济高效的获取任何 DNA 序列的方法，无论其大小或序列组成如何。它将让科学家专注于实验而不是剪切和粘贴 DNA 分子，从而加快研究步伐。它还将使新项目成为可能，如果不完全控制大型 DNA 序列，这些项目将是不可能或不切实际的昂贵的。