Studies of DNA Methyltransferases

DNA甲基转移酶的研究

• 批准号: 8639587
• 负责人: RICHARD J ROBERTS
• 金额: $ 50.11万
• 依托单位: NEW ENGLAND BIOLABS, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-28 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8639587
• 关键词: Base Sequence; Bioinformatics; DNA; DNA Methyltransferase; DNA Modification Methylases; DNA Restriction-Modification Enzymes; DNA Sequence; Detection; Engineering; Enzyme Gene; Enzymes; Family; Goals; Knowledge; Methylation; Methyltransferase; Methyltransferase Gene; New England; Phase; Plasmids; Positioning Attribute; Production; Property; Reagent; Research; Site; Specificity; System; Technology; Testing; Type II site-specific deoxyribonuclease; Work; base; gene discovery; genome annotation; improved; novel; pathogen; promoter; restriction enzyme

DESCRIPTION (provided by applicant): This project takes advantage of a DNA sequencing technology newly developed by Pacific Biosciences called SMRT sequencing. In addition to providing the DNA base sequence, this approach also allows the detection of modified bases. DNA methyltransferases have traditionally been rather difficult to characterize and as a result, we do not currently know the accurate DNA recognition specificity of any DNA methyltransferases. In the case of those that form part of restriction-modification systems, it has always been assumed that they will have the same recognition specificity as the restriction enzyme. However, in some systems preliminary results indicated that this might not be so, and we now in a position to test that explicitly. A knowledge of the specificity will be key on many fronts. Firstly, in some cases it will allow these methyltransferases to become valuable commercial reagents. Secondly, we expect to discover novel DNA methyltransferases with properties that would make them suitable for specific applications. One such enzyme discovered very recently, is M.EcoGI, which appears to recognize all A residues in a sequence and convert them at very high efficiency to N6- methyladenine. The results of this project will also greatly enhance the value of the cloned restriction-modification systems that we have at New England Biolabs and will also help considerably in the functional annotation of newly determined DNA sequences. It is an ideal blend of academic and commercial research and therefore is especially suited for New England Biolabs.

描述（由申请人提供）：该项目利用了由Pacific Biosciences新开发的称为SMRT测序的DNA测序技术。除了提供DNA碱基序列外，该方法还允许检测修饰的碱基。传统上，DNA甲基转移酶很难表征，因此，我们目前不知道任何DNA甲基转移酶的准确DNA识别特异性。对于那些构成限制修改系统一部分的人，它具有 始终认为它们将具有与限制酶相同的识别特异性。但是，在某些系统中，初步结果表明可能并非如此，现在我们可以明确测试。对特异性的了解将是许多方面的关键。首先，在某些情况下，这些甲基转移酶可以成为有价值的商业试剂。其次，我们希望发现具有特性的新型DNA甲基转移酶，使其适合特定应用。 M.Ecogi最近发现的一种这样的酶是序列中的所有A残基，并以非常高的效率将其转换为N6-甲基丹宁。该项目的结果还将大大提高我们在新英格兰Biolabs中拥有的克隆限制性修饰系统的价值，并且还将在新确定的DNA序列的功能注释中有很大帮助。它是学术和商业研究的理想融合，因此特别适合新英格兰生物群。