Fluorescent Assay for Directed Evolution of Perhydrolases
نویسندگان
چکیده
منابع مشابه
Two-photon directed evolution of green fluorescent proteins
Directed evolution has been used extensively to improve the properties of a variety of fluorescent proteins (FPs). Evolutionary strategies, however, have not yet been used to improve the two-photon absorption (2PA) properties of a fluorescent protein, properties that are important for two-photon imaging in living tissues, including the brain. Here we demonstrate a technique for quantitatively s...
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Microbial rhodopsins are a diverse group of photoactive transmembrane proteins found in all three domains of life. A member of this protein family, Archaerhodopsin-3 (Arch) of halobacterium Halorubrum sodomense, was recently shown to function as a fluorescent indicator of membrane potential when expressed in mammalian neurons. Arch fluorescence, however, is very dim and is not optimal for appli...
متن کاملDirected evolution of green fluorescent protein by a new versatile PCR strategy for site-directed and semi-random mutagenesis.
To develop a simple, speedy, economical and widely applicable method for multiple-site mutagenesis, we have substantially modified the Quik-Change Site-Directed Mutagenesis Kit protocol (Stratagene, La Jolla, CA). Our new protocol consists of (i) a PCR reaction using an in vitro technique, LDA (ligation-during-amplification), (ii) a DPN:I treatment to digest parental DNA and to make megaprimers...
متن کاملA flow cytometer-based whole cell screening toolbox for directed hydrolase evolution through fluorescent hydrogels.
A high throughput whole cell flow cytometer screening toolbox was developed and validated by identifying improved variants (1.3-7-fold) for three hydrolases (esterase, lipase, cellulase). The screening principle is based on coupled enzymatic reaction using glucose derivatives which yield upon hydrolysis a fluorescent-hydrogel-layer on the surface of E. coli cells.
متن کاملCorrection: A flow cytometer-based whole cell screening toolbox for directed hydrolase evolution through fluorescent hydrogels.
Correction for 'A flow cytometer-based whole cell screening toolbox for directed hydrolase evolution through fluorescent hydrogels' by Nina Lülsdorf et al., Chem. Commun., 2015, 51, 8679-8682.
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ژورنال
عنوان ژورنال: Journal of Biomolecular Screening
سال: 2012
ISSN: 1087-0571,1552-454X
DOI: 10.1177/1087057112438464