Abstract
The twin-arginine translocation (Tat) pathway is an attractive route for secretory production of heterologous proteins in E. coli. In this study, we investigated the potential use of Tat signal peptide from S. coelicolor to improve secretory expression. The results showed that Tat signal peptide (ssDagA) could effectively secrete active Green fluorescent protein (GFP) to periplasm. When the rare codons of signal sequence were optimized, the expression and secretion yield of GFP improved by about 2-3 folds as detected qualitatively by western blotting and fluorescent analysis. The increase of translation rate could be explained by the unstability of mRNA secondary structure. In summary, our strategy could provide a new approach for high-level secretory expression of heterologous proteins in E. coli.
Keywords: Twin-arginine translocation, codon usage, signal sequence, secretion
Protein & Peptide Letters
Title: New Approach to Achieve High-Level Secretory Expression of Heterologous Proteins by Using Tat Signal Peptide
Volume: 16 Issue: 6
Author(s): Yu-Dong Li, Zhan Zhou, Long-Xian Lv, Xiao-Ping Hou and Yong-Quan Li
Affiliation:
Keywords: Twin-arginine translocation, codon usage, signal sequence, secretion
Abstract: The twin-arginine translocation (Tat) pathway is an attractive route for secretory production of heterologous proteins in E. coli. In this study, we investigated the potential use of Tat signal peptide from S. coelicolor to improve secretory expression. The results showed that Tat signal peptide (ssDagA) could effectively secrete active Green fluorescent protein (GFP) to periplasm. When the rare codons of signal sequence were optimized, the expression and secretion yield of GFP improved by about 2-3 folds as detected qualitatively by western blotting and fluorescent analysis. The increase of translation rate could be explained by the unstability of mRNA secondary structure. In summary, our strategy could provide a new approach for high-level secretory expression of heterologous proteins in E. coli.
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Cite this article as:
Li Yu-Dong, Zhou Zhan, Lv Long-Xian, Hou Xiao-Ping and Li Yong-Quan, New Approach to Achieve High-Level Secretory Expression of Heterologous Proteins by Using Tat Signal Peptide, Protein & Peptide Letters 2009; 16 (6) . https://dx.doi.org/10.2174/092986609788490096
DOI https://dx.doi.org/10.2174/092986609788490096 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
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