Visualization of Periplasmic and Cytoplasmic Proteins with a Self-Labeling Protein Tag

Journal of Bacteriology
Na KeMehmet Berkmen

Abstract

The use of fluorescent and luminescent proteins in visualizing proteins has become a powerful tool in understanding molecular and cellular processes within living organisms. This success has resulted in an ever-increasing demand for new and more versatile protein-labeling tools that permit light-based detection of proteins within living cells. In this report, we present data supporting the use of the self-labeling HaloTag protein as a light-emitting reporter for protein fusions within the model prokaryote Escherichia coli. We show that functional protein fusions of the HaloTag can be detected both in vivo and in vitro when expressed within the cytoplasmic or periplasmic compartments of E. coli. The capacity to visually detect proteins localized in various prokaryotic compartments expands today's molecular biologist toolbox and paves the path to new applications. Visualizing proteins microscopically within living cells is important for understanding both the biology of cells and the role of proteins within living cells. Currently, the most common tool is green fluorescent protein (GFP). However, fluorescent proteins such as GFP have many limitations; therefore, the field of molecular biology is always in need of new tools to vis...Continue Reading

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Dec 4, 2019·Bioconjugate Chemistry·Harun F OzbakirArnab Mukherjee

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Methods Mentioned

BETA
fluorescence-activated cell sorter
PCR
motility assay
fluorescence microscopy

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