POWRE: Isolation and characterization of salt-sensitive mutants in the archaeal halophile, Haloferax volcanii

这一POWRE项目的重点是分离和鉴定嗜盐古细菌Haloferax Volcanii的盐敏感突变体。属于古生界的极端嗜盐细菌生长需要的盐浓度从-1.5M到5.2M氯化钠，这一上限超过了海水盐度的十倍。嗜盐生物在死海和大盐湖等恶劣环境中被发现，那里高浓度的盐阻止了几乎所有其他已知生物的生长。到目前为止，还没有文献报道使用基因技术，如突变，目的是在嗜盐古细菌中创造对盐敏感的突变。在这项研究中，将进行基因中断突变，以获得不再能够在3.5M氯化钠中生长的火山杆菌突变体，这是该生物的最佳盐浓度。将对突变体进行分析，以确定导致盐敏感表型的基因(S)的特定位置。负责盐敏感的基因(S)将被鉴定和鉴定。将对这些基因进行初步研究，以确定它们如何受到调节，以应对环境中的盐度。收集到的数据将导致未来研究极端嗜盐生物的基因调控。这项研究将为了解嗜盐微生物如何在体内从分子水平上适应高盐环境提供关键的第一步。此外，任何可以收集到的关于古生物用来对抗它们所居住的恶劣环境的影响的适应机制的遗传信息都是重要的。这个项目是探索性的，有可能在POWRE计划的目标下推进Bidle博士的职业生涯。

This POWRE project focuses on the isolation and characterization of salt-sensitive mutants of the halophilic Archaeon, Haloferax volcanii. The extreme halophiles belonging to the domain Archaea require salt concentrations for growth that range from -1.5 M up to 5.2 M NaCl, an upper limit in excess of ten times the salinity of seawater. Halophiles are found in such hostile environments as the Dead Sea and the Great Salt Lake, where high concentrations of salt preclude the growth of almost all other known organisms. To date, there are no documented reports that have used genetic techniques such as mutagenesis with the specific intent of creating salt-sensitive mutants in the halophilic Archaea. In this study, gene disruption mutagenesis will be performed to obtain H. volcanii mutants that are no longer capable of growing at 3.5 M NaCl, the optimal salt concentration for this organism. Mutants will be analyzed to determine the specific location of the gene(s) that was disrupted to cause the salt-sensitive phenotype. The gene(s) responsible for salt- sensitivity will be identified and characterized. Preliminary studies will be conducted on these genes to determine how they are regulated in response to salinity in the environment. The data gathered will lead to future studies examining gene regulation in the extreme halophiles. This study will provide the first crucial steps towards understanding how the halophiles are able to adapt in vivo to hypersaline environments on the molecular level. Furthermore, any genetic information that can be gathered about the adaptational mechanisms used by the Archaea to combat the effects of the harsh environments they inhabit is of importance. This project is of an exploratory nature and has the potential to advance Dr. Bidle's career under the objectives of the POWRE program.