Reddy, P.
and O'Dell, W.
(2021),
Fusing an Insoluble Protein to GroEL Apical Domain Enhances Soluble Expression in Escherichia coli, Methods in Enzymology Volume 659: Recombinant Protein Expression: Prokaryotic hosts and cell free systems, Elsevier, Philadelphia, PA, [online], https://doi.org/10.1016/bs.mie.2021.09.002, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=933220
(Accessed November 21, 2024)
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
Secure .gov websites use HTTPS
A lock (
) or https:// means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.
Fusing an Insoluble Protein to GroEL Apical Domain Enhances Soluble Expression in Escherichia coli
Published
Author(s)
Prasad 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 were cloned into the pRE:GroEL fusion vector and successfully expressed as fusion proteins in Escherichia coli.Citation
Methods in Enzymology Volume 659: Recombinant Protein Expression: Prokaryotic hosts and cell free systems
Volume
659
Publisher Info
Elsevier, Philadelphia, PA
Pub Type
Book Chapters
Download Paper
Keywords
pRE recombinant plasmid, GroEL minichaperone, fusion protein expression, soluble protein production
Citation
Issues
If you have any questions about this publication or are having problems accessing it, please contact reflib@nist.gov.
Created October 29, 2021, Updated November 29, 2022