Davis Chris R, Johnson Carl H, Robertson J Brian
Department of Biology, Middle Tennessee State University, Murfreesboro, TN, USA.
Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA.
Extremophiles. 2020 Sep;24(5):773-785. doi: 10.1007/s00792-020-01193-x. Epub 2020 Aug 4.
Haloarchaea have evolved to thrive in hypersaline environments. Haloferax volcanii is of particular interest due to its genetic tractability; however, few in vivo reporters exist for halophiles. Haloarchaeal proteins evolved characteristics that promote proper folding and function at high salt concentrations, but many mesophilic reporter proteins lack these characteristics. Mesophilic proteins that acquire salt-stabilizing mutations, however, can lead to proper function in haloarchaea. Using laboratory-directed evolution, we developed and demonstrated an in vivo luciferase that functions in the hypersaline cytosol of H. volcanii.
嗜盐古菌已经进化到能够在高盐环境中茁壮成长。沃氏嗜盐富球菌因其遗传易处理性而特别受关注;然而,嗜盐菌的体内报告基因很少。嗜盐古菌蛋白质进化出了在高盐浓度下促进正确折叠和发挥功能的特性,但许多嗜温报告蛋白缺乏这些特性。然而,获得盐稳定突变的嗜温蛋白可以在嗜盐古菌中发挥正常功能。通过实验室定向进化,我们开发并证明了一种能在沃氏嗜盐富球菌的高盐细胞质中发挥作用的体内荧光素酶。