Linking any protein to GroEL minichaperone enhances protein production and solubility in Escherichia coli.

Published

September 23, 2021

Author(s)

Prasad T. Reddy

Abstract

A protocol for increasing soluble protein expression by fusing the chaperone GroEL apical domain with a gene of interest is described herein. GroEL apical domain, the minichaperone that functions independently of GroES and ATP in protein folding, is cloned downstream of the lambda CII ribosome binding site in the parent pRE vector. The pRE vector has tightly controlled transcription suitable for expressing toxic proteins. The GroEL minichaperone is fused to a glycine-serine rich linker followed by the enterokinase protease recognition sequence. A number of genes that are recalcitrant to protein production in the parent pRE vector 5were cloned into the pRE:GroEL fusion vector and successfully expressed as fusion proteins in Escherichia coli.

Citation

Methods in Enzymology: Recombinant Protein Expression: Prokaryotic Hosts and Cell-Free Systems

Volume

659

Publisher Info

Elsevier, Amsterdam,

Pub Type

Book Chapters

Download Paper

https://doi.org/10.1016/bs.mie.2021.09.002

Local Download

Keywords

Plasmid vector, Protein production

Citation

Reddy, P. (2021), Linking any protein to GroEL minichaperone enhances protein production and solubility in Escherichia coli., Methods in Enzymology: Recombinant Protein Expression: Prokaryotic Hosts and Cell-Free Systems, Elsevier, Amsterdam, , [online], https://doi.org/10.1016/bs.mie.2021.09.002, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=922005 (Accessed October 14, 2025)