Directed Evolution of Streptavidin Variants Using In Vitro Compartmentalization
نویسندگان
چکیده
منابع مشابه
Directed evolution of streptavidin variants using in vitro compartmentalization.
We have developed and implemented an in vitro compartmentalization (IVC) selection scheme for the identification of streptavidin (SA) variants with altered specificities for the biotin analog desthiobiotin. Wild-type SA and selected variants bind desthiobiotin with similar affinities (approximately 10(-13) M), but the variants have off rates almost 50 times slower and a half-life for dissociati...
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Engineering the specificity of DNA-modifying enzymes has proven extremely challenging, as sequence recognition by these enzymes is poorly understood. Here we used directed evolution to generate a variant of HaeIII methyltransferase that efficiently methylates a novel target site. M.HaeIII methylates the internal cytosine of the canonical sequence GGCC, but there is promiscuous methylation of a ...
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ژورنال
عنوان ژورنال: Chemistry & Biology
سال: 2008
ISSN: 1074-5521
DOI: 10.1016/j.chembiol.2008.07.017