Next Generation DNA Synthesis

下一代 DNA 合成

• 批准号: BB/M025624/1
• 负责人: Tom Brown
• 金额: $ 282.74万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FM025624%2F1
• 关键词: Next; Generation; DNA; Synthesis

Ever large pieces of DNA such as genes and gene clusters are required for Synthetic Biology, and these are normally made by a combination of chemical and biochemical methods. The chemical methodology is required at the start of the process to generate very short pieces of DNA (oligonucleotides) by automated solid-phase methods. These are then used to build bigger pieces of DNA by biochemical methods that are based on the polymerase chain reaction (PCR amplification). The chemical synthesis of DNA can lead to damage which results in mistakes (mutations) in the final DNA product, and to avoid this the DNA has to be repaired by various enzymes. This is tedious and slows down the overall process, increasing costs and limiting the size of DNA that can be made. In this project we will analyse DNA made by modern ultra high throughput chemical methods and optimise the process to minimise mutations. We will also explore a different way to make large pieces of DNA; enzymatic ligation. In this process DNA constructs with modified bases can be made, which are very useful in gene expression and biomedical studies. These cannot be made by PCR amplification which erazes the modifications. Such modified DNA can only be properly made from highly pure oligonucleotides in very large numbers, placing stringent requiremenst on high-throughput oligonucleotide synthesis. Overall this project will greatly increase the capacity, quality and efficiency of DNA synthesis and is highly relevant to Synthetic Biology Centres in the UK and beyond.

合成生物学需要更大的DNA片段，如基因和基因簇，这些通常是通过化学和生物化学方法的结合来制造的。化学方法需要在过程开始时通过自动固相法生成非常短的DNA片段（寡核苷酸）。然后用基于聚合酶链反应（PCR扩增）的生化方法来构建更大的DNA片段。DNA的化学合成会导致损伤，从而导致最终的DNA产物出现错误（突变），为了避免这种情况，DNA必须通过各种酶进行修复。这是乏味的，减缓了整个过程，增加了成本，限制了DNA的大小。在这个项目中，我们将分析由现代超高通量化学方法制造的DNA，并优化过程以最大限度地减少突变。我们还将探索另一种制造大片段DNA的方法；酶结扎。在这一过程中，可以构建带有修饰碱基的DNA结构，这在基因表达和生物医学研究中非常有用。这些不能通过PCR扩增产生，因为扩增会抹去修饰。这种修饰的DNA只能由非常大量的高纯度寡核苷酸正确合成，这对高通量寡核苷酸合成提出了严格的要求。总体而言，该项目将大大提高DNA合成的能力、质量和效率，并与英国及其他地区的合成生物学中心高度相关。

项目成果

Expression and In Vivo Loading of De Novo Proteins with Tetrapyrrole Cofactors.

使用四吡咯辅因子表达和体内装载 De Novo 蛋白质。

• DOI: 10.1007/978-1-0716-1826-4_8
• 发表时间: 2022
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Curnow P

A New Micromonospora Strain with Antibiotic Activity Isolated from the Microbiome of a Mid-Atlantic Deep-Sea Sponge.

• DOI: 10.3390/md19020105
• 发表时间: 2021-02-11
• 期刊: Marine drugs
• 影响因子: 5.4
• 作者: Back CR;Stennett HL;Williams SE;Wang L;Ojeda Gomez J;Abdulle OM;Duffy T;Neal C;Mantell J;Jepson MA;Hendry KR;Powell D;Stach JEM;Essex-Lopresti AE;Willis CL;Curnow P;Race PR
• 通讯作者: Race PR

Single Site Discrimination of Cytosine, 5-Methylcytosine, and 5-Hydroxymethylcytosine in Target DNA Using Anthracene-Tagged Fluorescent Probes

使用蒽标记荧光探针对目标 DNA 中的胞嘧啶、5-甲基胞嘧啶和 5-羟甲基胞嘧啶进行单位点鉴别

• DOI: 10.1021/acschembio.5b00796
• 发表时间: 2015
• 期刊: ACS Chemical Biology
• 影响因子: 4
• 作者: Duprey J

Leaf LIMS: A Flexible Laboratory Information Management System with a Synthetic Biology Focus.

Leaf LIMS：以合成生物学为重点的灵活实验室信息管理系统。

• DOI: 10.1021/acssynbio.7b00212
• 发表时间: 2017
• 期刊: ACS synthetic biology
• 影响因子: 4.7
• 作者: Craig T