HaloTag7: A genetically engineered tag that enhances bacterial expression of soluble proteins and improves protein purification

Ohana, Rachel Friedman; Encell, Lance P.; Zhao, Kate; Simpson, Dan; Slater, Michael; Urh, Marjeta; Wood, Keith V.

doi:10.1016/j.pep.2009.05.010

Cited by 140 publications

(126 citation statements)

References 42 publications

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“…In the 58 course of this work, we observed that the N-terminal HaloTag fusion had a substantial effect 59 on the expression of the more recalcitrant FGFs, consistent with the observation that HaloTag 60 is a potential solubilisation tag for recombinant proteins (Ohana et al 2009). Thus, whereas 61 expression of FGF1 and FGF10 was somewhat reduced and that of FGF2 increased, 62 expression of FGF7, which can be toxic (Ron et al 1993) was no longer so, while expression 63 of soluble FGF3, FGF6, FGF7, FGF8, FGF16, FGF17, FGF20 and FGF22 was markedly 64 enhanced.…”

supporting

confidence: 73%

HaloTag is an effective expression and solubilisation fusion partner for a range of fibroblast growth factors

Sun

Taylor

et al. 2014

Preprint

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supporting

confidence: 73%

HaloTag is an effective expression and solubilisation fusion partner for a range of fibroblast growth factors

Sun

Taylor

et al. 2014

Preprint

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“…HLDs catalyze reactions of great environmental and biotechnological significance. They have potential applications in bioremediation (33), biosensing (4,6), decontamination of warfare agents (30), synthesis of optically pure compounds (29,35), cellular imaging, and protein analysis (25,27). The effective use of biocatalysts in these applications requires availability of enzymes with specific properties under process conditions (5).…”

mentioning

confidence: 99%

Biochemical Characterization of a Novel Haloalkane Dehalogenase from a Cold-Adapted Bacterium

Drienovská

Chovancová

Koudeláková

et al. 2012

Appl Environ Microbiol

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ABSTRACTA haloalkane dehalogenase, DpcA, fromPsychrobacter cryohalolentisK5, representing a novel psychrophilic member of the haloalkane dehalogenase family, was identified and biochemically characterized. DpcA exhibited a unique temperature profile with exceptionally high activities at low temperatures. The psychrophilic properties of DpcA make this enzyme promising for various environmental applications.

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“…Moreover, when compared with fluorescein-labeled systems, the eosin-labeled system exhibits a fivefold greater efficiency in ROS generation, which was measured using anthracene-9,10-dipropionic acid (Takemoto et al, 2013). Eosin is also used in combination with a protein tag, HaloTag7, a haloalkane dehalogenase mutant (Ohana et al, 2009), which has been fused to protein kinase Cc (PKCc). Upon the chemical stimulation, PKCc translocates to the plasma membrane.…”

Section: Cali With Transgenically Encoded Tagsmentioning

confidence: 99%

Chromophore-assisted laser inactivation – towards a spatiotemporal–functional analysis of proteins, and the ablation of chromatin, organelle and cell function

Sano

Watanabe

Matsunaga

2014

Journal of Cell Science

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Chromophore-assisted laser or light inactivation (CALI) has been employed as a promising technique to achieve spatiotemporal knockdown or loss-of-function of target molecules in situ. CALI is performed using photosensitizers as generators of reactive oxygen species (ROS). There are two CALI approaches that use either transgenic tags with chemical photosensitizers, or genetically encoded fluorescent protein fusions. Using spatially restricted microscopy illumination, CALI can address questions regarding, for example, protein isoforms, subcellular localization or phase-specific analyses of multifunctional proteins that other knockdown approaches, such as RNA interference or treatment with chemicals, cannot. Furthermore, rescue experiments can clarify the phenotypic capabilities of CALI after the depletion of endogenous targets. CALI can also provide information about individual events that are involved in the function of a target protein and highlight them in multifactorial events. Beyond functional analysis of proteins, CALI of nuclear proteins can be performed to induce cell cycle arrest, chromatin-or locus-specific DNA damage. Even at organelle levelsuch as in mitochondria, the plasma membrane or lysosomes -CALI can trigger cell death. Moreover, CALI has emerged as an optogenetic tool to switch off signaling pathways, including the optical depletion of individual neurons. In this Commentary, we review recent applications of CALI and discuss the utility and effective use of CALI to address open questions in cell biology.

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HaloTag7: A genetically engineered tag that enhances bacterial expression of soluble proteins and improves protein purification

Cited by 140 publications

References 42 publications

HaloTag is an effective expression and solubilisation fusion partner for a range of fibroblast growth factors

HaloTag is an effective expression and solubilisation fusion partner for a range of fibroblast growth factors

Biochemical Characterization of a Novel Haloalkane Dehalogenase from a Cold-Adapted Bacterium

Chromophore-assisted laser inactivation – towards a spatiotemporal–functional analysis of proteins, and the ablation of chromatin, organelle and cell function

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