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
(Accessed May 7, 2024)
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Linking any protein to GroEL minichaperone enhances protein production and solubility in Escherichia coli.
Published
Author(s)
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
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Keywords
Plasmid vector, Protein production
Citation
Created September 23, 2021, Updated October 11, 2022