A Model Archaeal System: Genetic Tools and Resources for T. kodakarensis

模型古菌系统：T. kodakarensis 的遗传工具和资源

• 批准号: 8322564
• 负责人: JOHN NEWTON REEVE
• 金额: $ 29.92万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8322564
• 关键词: Affinity; Affinity Chromatography; Amino Acids; Animal Model; Archaea; Biological; Biological Models; Biology; Biomedical Research; Cell Density; Cell division; Cells; Communities; Complex; Cytokinesis; DNA Repair; DNA biosynthesis; Development; Disease; Epitopes; Essential Genes; Eukaryota; Eukaryotic Cell; Experimental Models; Gene Expression; Gene Library; Gene Targeting; Genes; Genetic; Genetic Information Processing Pathway; Genetic Techniques; Genetic Transcription; Genome; Genomics; Health; Health Services Research; Hemagglutinin; Histidine; Homologous Gene; Human; Human Biology; Internet; Investigation; Laboratories; Libraries; Liquid substance; Mass Spectrum Analysis; Modeling; Molecular; Molecular Biology; Molecular and Cellular Biology; One-Step dentin bonding system; Open Reading Frames; Palpable; Plasmids; Procedures; Process; Prokaryotic Cells; Proteins; Recombinants; Regulation; Relative (related person); Research; Research Personnel; Resources; System; Thermococcus; Transcript; Translation Initiation; Work; genetic resource; genome sequencing; in vivo; knockout gene; programs; protein expression; research study; systems research; tool; vector

DESCRIPTION (provided by applicant): Research on bacterial nnodel systems has established much of our understanding of basic molecular biology. Extrapolations from bacterial systems to eukaryotic molecular biology and human health issues are however inherently limited by the lack of bacterial homologues of many components conserved in eukaryotic cells. Fortunately, Archaea do have homologues of many of these "eukaryotic" components and research with such simpler archaeal systems can therefore be legitimately extrapolated to eukaryotic/human cellular and molecular biology. The archaeal machineries, for example, that catalyze DNA replication and repair, transcription, transcript and protein processing, translation initiation, cytokinesis and cell division are all far simpler than their eukaryotic counterparts, but most of their components are proteins that have well-conserved structural and likely functional eukaryotic homologues. For experimental research, the attraction of these much simpler but eukaryote-homologous prokaryotic systems is obvious and palpable, but with very limited genetics and no archaeal model system, access of the research community to the advantages and opportunities afforded by the Archaea has been effectively blocked. We have recently established the genetic techniques needed for Thermococcus kodakarensis (T.k.) and, as solicited by the PA-07-457 request "...to develop and distribute genetic and genomic resources for emerging non-mammalian model organisms" the project proposed will generate the genomic resources needed to fully establish T.k. as a facile archaeal model system. Specifically, every protein-encoding gene will be Individually cloned and also modified to add a hemagglutinin (HA) epitope for Identification and a Hls6-afflnlty-tag for purification In sequence-verified plasmid libraries. T.k. strain libraries will also be constructed with every non-essential open reading frame (ORF) Individually deleted, and every genomic ORF HA-His6 extended. With the T.k. genetic techniques established and these genomic resources constructed, the first archaeal model system will be available. This will facilitate, expedite and encourage Investigations ofthe many archaeal cellular and molecular biology svstems that are homologues of eukarvotic/human systems and so valid models for healthcare research.

描述（由申请人提供）：细菌NNODEL系统的研究已经确定了我们对基本分子生物学的大部分理解。但是，从细菌系统到真核分子生物学和人类健康问题的外推受到了本质上受到真核细胞中许多成分的细菌同源物的限制。幸运的是，古细菌确实具有许多这些“真核”成分的同源物，因此，使用这种简单的古细菌系统的研究可以合法地外推到真核/人类细胞和分子生物学。例如，古老的机器可以催化DNA复制和修复，转录，转录和蛋白质加工，翻译起始，细胞因子和细胞分裂都比其真核对应物更简单，但是它们的大多数成分都是蛋白质，具有良好的结构性和可能功能性的真主性真菌性化合物。对于实验研究，这些更简单但真核的原始原核系统的吸引力是显而易见且可显着的，但是由于遗传学非常有限，而没有古细菌模型系统，因此研究界的访问权限已被有效地阻止古细菌所提供的优势和机会。我们最近已经建立了热门柯达卡菌（T.K.）所需的遗传技术，并由PA-07-457请求征求的那样，开发和分发了新兴的非乳腺模型生物体的遗传和基因组资源”，该项目提出的项目将产生完全建立T.K的基因组资源。作为简便的古细菌模型系统。具体而言，每个蛋白质编码的基因都将被单独克隆并修改，以添加血凝素（HA）表位，以识别识别和HLS6-AFFLNLTY-TAG，以在序列验证的质粒文库中纯化。 T.K.应变库也将使用每个非必需的开放阅读框（ORF）单独删除，并且每个基因组ORF HA-HIS6延伸。与T.K.建立的遗传技术以及建立的这些基因组资源，将提供第一个古细菌模型系统。这将促进，加快和鼓励对许多是Eukarvitotic/Human Systems的同源物以及医疗保健研究的有效模型的许多古细胞和分子生物学系统的研究。