FINDING NOVEL REAGENTS BY BIOINFORMATICS AND BIOCHEMISTRY

通过生物信息学和生物化学寻找新试剂

• 批准号: 8488588
• 负责人: RICHARD J ROBERTS
• 金额: $ 43.74万
• 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: 2012-03-05 至 2014-09-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8488588
• 关键词: Archaea; Biochemical; Biochemistry; Bioinformatics; Burkholderia; Cells; Cleaved cell; Clinical; DNA; DNA Methyltransferase; DNA Modification Methylases; DNA Restriction Enzymes; DNA Restriction-Modification Enzymes; Development; Engineering; Enzymes; Epigenetic Process; Genbank; Gene Family; Genes; Genetic; Genome; Homologous Gene; Knock-out; Methods; Methyltransferase; Methyltransferase Gene; Modification; Molecular Biology; Nature; Nucleosides; Organism; Pathogenesis; Phase; Preparation; Property; Reagent; Research; Research Personnel; Screening procedure; Shotgun Sequencing; Specificity; Speed; Streptomyces coelicolor; System; Techniques; Type II site-specific deoxyribonuclease; Work; base; cost; genome sequencing; member; novel; pathogen; pathogenic bacteria; phosphorothioate; programs; restriction enzyme; tool

DESCRIPTION (provided by applicant): This project aims to find novel restriction enzymes that will be useful in many aspects of molecular biology research. One particular focus will be on restriction endonucleases that are able to recognize modified DNA, especially DNA containing 5-methylcytosine and 5- hydroxymethycytosine since such enzymes have many potential applications in epigenetics research. Enzymes recognizing these modified bases are known to occur in bacterial and archaea strains and in the past, many have been found by random biochemical screening. In the proposed project, the discovery of potential new enzymes will use bioinformatics techniques and take advantage of the large number of currently sequenced prokaryotic genomes, both completely sequenced genome and shotgun sequences. The project will identify good candidates and then use biochemical methods to characterize them. In addition to commercializing any useful enzymes that are found from this screen, the information gained about the biochemical function of the restriction enzymes discovered will be invaluable for genome annotation and will have practical implications for researchers working with pathogenic organisms. The latter will result from our efforts to characterize the restriction-modification systems in pathogens and identify those systems which need either to be knocked out or circumvented by prior modification prior to transformation. Thus, the results of this research will have immediate implications for the fields of epigenetics, pathogenesis and genome annotation.

描述（由申请人提供）：该项目旨在找到新颖的限制酶，这些酶在分子生物学研究的许多方面都有用。一个特别的重点将是能够识别改良的DNA的限制性核酸内切酶，尤其是含有5-甲基胞嘧啶和5-羟基羟基胞霉素的DNA，因为这种酶在表观遗传学研究中具有许多潜在的应用。已知识别这些修饰碱基的酶发生在细菌和古细菌菌株中，过去，许多通过随机生化筛选发现了许多酶。在拟议的项目中，潜在的新酶的发现将使用生物信息学技术，并利用当前测序的原核生物基因组的大量，都是完全测序的基因组和shot弹枪序列。该项目将确定良好的候选人，然后使用生化方法来表征它们。除了从此屏幕上发现的任何有用的酶进行商业化外，还获得有关发现的限制酶的生化功能的信息对于基因组注释将是无价的，并且对使用致病生物的研究人员具有实际意义。后者将源于我们为病原体中的限制性修饰系统表征并确定需要在转换之前先进行先验修改而绕开的系统。因此，这项研究的结果将对表观遗传学，发病机理和基因组注释的领域具有直接的影响。