ISSN:
1432-1432
Keywords:
In vitro selection
;
Bacteriophage evolution
;
Protein stability
;
Urea
;
Extremophile
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Extremophiles are microorganisms that thrive under extreme conditions such as temperatures above 65°C, pHs below 4 or above 10, salt concentrations above 0.5 m, or pressures of 600 atm. While studies of enzymes either isolated from extremophiles, or generated using site-specific mutagenesis, or adapted by in vivo or in vitro selection have established a precedent for the engineering and application of proteins at extreme conditions, generalization of the approaches to more complex multimolecular or multitask systems has remained elusive. Here we demonstrate that a significantly more complex system—a bacteriophage—can over a number of generations be adapted to tolerate a hostile and unnatural environment. An in vitro selection strategy was used to adapt phage to urea, a protein denaturing agent. As the concentration of urea employed in selections over 20 generations was gradually increased from 5 to 9 m, the surviving phages steadily improved their tolerance, finally achieving a greater than 350-fold stability enhancement over the original population.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00170661
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