Expressing red fluorescent protein on the surface of Escherichia coli using C-terminal domain of autotransporters

Document Type : Short communication

Authors

1 Department of Molecular and Environmental Biotechnology, Faculty of Biology and Biotechnology, University of Science, Ho Chi Minh city, Vietnam

2 Vietnam National University, Ho Chi Minh city, Vietnam

3 Labolatory of Biosensors, Faculty of Biology and Biotechnology, University of Science, Ho Chi Minh city, Vietnam

Abstract

The Type V secretion system, or “autotransporter”, is a secretion system that enables bacteria to directly export proteins from the cell interior to the extracellular membrane. mCherry is a second-generation monomeric red fluorescent protein that has an improvement in photostability compared to the first generation of RFP. In this research, we conducted the fusion of the mRFP into the C-terminal domain of EhaA – the translocation domain of the autotransporter protein transport system – to investigate the expression of mRFP on the surface of Escherichia coli, a model organism commonly utilized in recombinant protein research. The induction of the mRFP-EhaA C-terminal domain complex expression was achieved using isopropyl β-D-1-thiogalactopyranoside (IPTG) and confirmed through SDS-PAGE stained with Coomassie Brilliant Blue and Western blotting using anti-6X His tag antibodies. The surface expression of the mRFP-EhaA C-terminal complex protein was validated through the fluorescent properties of mRFP and further confirmed using fluorescent microscopy. This study laid the groundwork for surface expression on cost-effective Gram-negative bacteria, E. coli.

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