Preparation and extraction of insoluble (inclusion-body) proteins from Escherichia coli.

Current Protocols in Protein Science
Ira Palmer, Paul T Wingfield

Abstract

High-level expression of many recombinant proteins in Escherichia coli leads to the formation of highly aggregated protein commonly referred to as inclusion bodies. Inclusion bodies are normally formed in the cytoplasm; alternatively, if a secretion vector is used, they can form in the periplasmic space. Inclusion bodies can be recovered from cell lysates and this unit describes preparation of washed pellets and solubilization of the protein using guanidine x HCl. The extracted protein, which is unfolded, is either directly folded as described in UNIT or further purified by gel filtration in the presence of guanidine x HCl as idescribed here. A support protocol describes the removal of guanidine x HCl from column fractions so they can be monitored by SDS-PAGE.

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Citations

Oct 27, 2017·The Journal of Immunology : Official Journal of the American Association of Immunologists·Adeola A ObajemuOlusegun O Onabajo
Nov 11, 2018·Bioscience Reports·Lalith K ChagantiKakoli Bose
Jan 24, 2009·Protein Science : a Publication of the Protein Society·Elizabeth Massey-GendelJames U Bowie
Mar 4, 2017·Scientific Reports·Izzat Fahimuddin Bin Mohamed SuffianKhuloud T Al-Jamal
Sep 5, 2008·Current Protocols in Microbiology·Wenping Qiu, Herman B Scholthof
Jul 24, 2018·Journal of Chemical Technology and Biotechnology·Vishwanath HebbiAnurag S Rathore
Apr 9, 2018·Applied Microbiology and Biotechnology·Ruramayi M NzumaIrene R Grant
Jul 6, 2019·Frontiers in Microbiology·Guillem Prats-EjarqueEster Boix
Feb 28, 2019·The Journal of Cell Biology·Hanaa HaririW Mike Henne
Jan 19, 2012·Microbial Cell Factories·Bihong ZhouZhanglin Lin
May 10, 2011·Protein Expression and Purification·Richard G DiScipioIngrid U Schraufstatter

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