Abstract
In prokaryotes, the twin-arginine translocation (Tat) signal peptide is specific for the secretion of folded proteins. On the other hand, the enhanced green fluorescent protein (EGFP) is widely used as a reporter protein in biological studies. In this work, the plasmids pBT2-ET-EGFP (encoding EGFP gene without signal peptide) and the plasmid pBT2-ETG (encoding the Tat-EGFP fusion protein with a staphylococcal TAT signal peptide) were respectively employed to investigate the translocation effect of Tat-EGFP by Tat signal peptide from the cytoplasm to periplasm of E. coli host. By examinations with microscopic fluorescence, SDS-PAGE and Western blot, as expected, the Tat-EGFP was successfully translocated into the periplasm of the E. coli host cells, but EGFP without signal peptide not. Furthermore, both kinds of EGFPs were not secreted into the fermentation broth. Therefore, this result primarily revealed the insight into the role of the heterologous signal peptide for the transmembrane translocation in E. coli.
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Acknowledgements
This work was financially supported by the National Natural Science Foundation of China (31370075, 31101275 and 61603273), the Undergraduate Laboratory Innovation Fund of Tianjin University of Science and Technology of China (1504A304X) and the Youth Innovation Fund of Tianjin University of Science and Technology of China (2014CXLG28).
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Zhou, QX., Zhang, J., Wang, MN., Yang, WH., Zhang, J., Gao, Q. (2018). Study on a Staphylococcal Tat Signal Peptide Guided EGFP Translocation in E. coli . In: Liu, H., Song, C., Ram, A. (eds) Advances in Applied Biotechnology. ICAB 2016. Lecture Notes in Electrical Engineering, vol 444. Springer, Singapore. https://doi.org/10.1007/978-981-10-4801-2_9
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DOI: https://doi.org/10.1007/978-981-10-4801-2_9
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