Hileman Travis H, Santangelo Thomas J
Department of Microbiology, Center for RNA Biology, Ohio State University Columbus, OH, USA.
Front Microbiol. 2012 Jun 8;3:195. doi: 10.3389/fmicb.2012.00195. eCollection 2012.
Thermococcus kodakarensis (T. kodakarensis) has emerged as a premier model system for studies of archaeal biochemistry, genetics, and hyperthermophily. This prominence is derived largely from the natural competence of T. kodakarensis and the comprehensive, rapid, and facile techniques available for manipulation of the T. kodakarensis genome. These genetic capacities are complemented by robust planktonic growth, simple selections, and screens, defined in vitro transcription and translation systems, replicative expression plasmids, in vivo reporter constructs, and an ever-expanding knowledge of the regulatory mechanisms underlying T. kodakarensis metabolism. Here we review the existing techniques for genetic and biochemical manipulation of T. kodakarensis. We also introduce a universal platform to generate the first comprehensive deletion and epitope/affinity tagged archaeal strain libraries.
嗜热栖热菌（Thermococcus kodakarensis，简称T. kodakarensis）已成为研究古细菌生物化学、遗传学和嗜热特性的首要模式系统。这种突出地位很大程度上源于嗜热栖热菌的天然感受态以及可用于操作嗜热栖热菌基因组的全面、快速且简便的技术。这些遗传能力辅以强劲的浮游生长、简单的筛选和选择方法、体外转录和翻译系统、复制型表达质粒、体内报告构建体以及对嗜热栖热菌代谢潜在调控机制不断扩展的认识。在此，我们综述了现有用于嗜热栖热菌遗传和生化操作的技术。我们还介绍了一个通用平台，用于构建首个全面的缺失和表位/亲和标签古细菌菌株文库。
