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Le-Hoang, Khoi-Nguyen, Nguyen, Thanh-Tan, Tran-Van, Hieu. (1403). Expressing red fluorescent protein on the surface of Escherichia coli using C-terminal domain of autotransporters. , 14(1), 31-35. doi: 10.22099/mbrc.2024.49860.1956
Khoi-Nguyen Le-Hoang; Thanh-Tan Nguyen; Hieu Tran-Van. "Expressing red fluorescent protein on the surface of Escherichia coli using C-terminal domain of autotransporters". , 14, 1, 1403, 31-35. doi: 10.22099/mbrc.2024.49860.1956
Le-Hoang, Khoi-Nguyen, Nguyen, Thanh-Tan, Tran-Van, Hieu. (1403). 'Expressing red fluorescent protein on the surface of Escherichia coli using C-terminal domain of autotransporters', , 14(1), pp. 31-35. doi: 10.22099/mbrc.2024.49860.1956
Le-Hoang, Khoi-Nguyen, Nguyen, Thanh-Tan, Tran-Van, Hieu. Expressing red fluorescent protein on the surface of Escherichia coli using C-terminal domain of autotransporters. , 1403; 14(1): 31-35. doi: 10.22099/mbrc.2024.49860.1956

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

Molecular Biology Research Communications
دوره 14، شماره 1، 2025، صفحه 31-35    اصل مقاله (176.04 K)
نوع مقاله: Short communication
شناسه دیجیتال (DOI): 10.22099/mbrc.2024.49860.1956
نویسندگان
Khoi-Nguyen Le-Hoang1، 2؛ Thanh-Tan Nguyen3؛ Hieu Tran-Van* 2
1Department of Molecular and Environmental Biotechnology, Faculty of Biology and Biotechnology, University of Science, Ho Chi Minh city, Vietnam
2Vietnam National University, Ho Chi Minh city, Vietnam
3Labolatory of Biosensors, Faculty of Biology and Biotechnology, University of Science, Ho Chi Minh city, Vietnam
چکیده
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.
کلیدواژه‌ها
Autotransporter؛ mRFP؛ C-terminal of EhaA؛ mRFP-C-terminal complex protein
مراجع
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