New approach to achieve high-level secretory expression of heterologous proteins by using Tat signal peptide.
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.
Li
Yu-Dong
YD
Zhejiang University, College of Life Sciences, Hangzhou, PR China.
Zhou
Zhan
Z
Lv
Long-Xian
LX
Hou
Xiao-Ping
XP
Li
Yong-Quan
YQ
D001426:Bacterial Proteins / Q000235:genetics*; D015153:Blotting, Western; D003062:Codon; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000235:genetics*; D009690:Nucleic Acid Conformation; D021382:Protein Sorting Signals / Q000235:genetics*; D012333:RNA, Messenger / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D048372:Streptomyces coelicolor / Q000235:genetics
D001426:Bacterial Proteins; D003062:Codon; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins