Publication. SBC

Title:

Translocation of green fluorescent protein by comparative analysis with multiple signal peptides

Author:

Linton, E , Walsh, M.K., Sims, R.C. and Miller, C.D.

Journal:

Biotechnology Journal

Abstract:

Type I and II secretory pathways are used for the translocation of recombinant proteins from the cytoplasm of Escherichia coli. The purpose of this study was to evaluate four signal peptides (HlyA, TorA, GeneIII, and PelB), representing the most common secretion pathways in E. coli, for their ability to target green fluorescent protein (GFP) for membrane translocation. Signal peptide-GFP genetic fusions were designed in accordance with BioFusion standards (BBF RFC 10, BBF RFC 23). The HlyA signal peptide targeted GFP for secretion to the extracellular media via the type I secretory pathway, whereas TAT-dependent signal peptide TorA and Sec-dependent signal peptide GeneIII exported GFP to the periplasm. The PelB signal peptide was inefficient in translocating GFP. The use of biological technical standards simplified the design and construction of functional signal peptide-recombinant protein genetic devices for type I and II secretion in E. coli. The utility of the standardized parts model is further illustrated as constructed biological parts are available for direct application to other studies on recombinant protein translocation.

Citation:

Linton, E , Walsh, M.K., Sims, R.C. and Miller, C.D. 2012. Translocation of green fluorescent protein by comparative analysis with multiple signal peptides. Biotechnology Journal. 7(5):667-76 doi 10.1002/biot.201100158. (0) IF:2.970

Hyperlink:

http://www.ncbi.nlm.nih.gov/pubmed/21834133