Recombinant protein purification by self-cleaving elastin-like polypeptide fusion tag

Current Protocols in Protein Science
Wan-Yi WuDavid W Wood

Abstract

This unit presents a rapid and simple method for the nonchromatographic purification of recombinant proteins expressed in E. coli. This method relies on a thermally responsive elastin-like polypeptide (ELP) tag, where the tagged protein is precipitated using a mild temperature shift. The tag is then induced to self-cleave by a mild pH shift and is subsequently removed by a final thermal precipitation. The result is a purified native protein target, without the requirement for affinity apparatus or protease removal of the tag. This protocol describes the required cloning methods to insert a given target into the expression vector, as well as the general method for purifying the resulting expressed protein.

References

Sep 3, 1999·Nature Biotechnology·D W WoodM Belfort
Nov 28, 2000·Annual Review of Genetics·X Q Liu
Nov 24, 2004·Protein Science : a Publication of the Protein Society·Kimberly Trabbic-CarlsonAshutosh Chilkoti
Aug 3, 2005·Nature Methods·Mahmoud Reza BankiDavid W Wood
Aug 11, 2005·Journal of the American Chemical Society·Xin GeCarlos D M Filipe
Nov 15, 2005·Microbial Cell Factories·Mahmoud Reza Banki, David W Wood
Nov 18, 2005·Journal of Molecular Biology·Kaori HiragaMarlene Belfort
Aug 30, 2008·Biotechnology and Bioengineering·Alison R GilliesDavid W Wood
Apr 7, 2009·Protein Expression and Purification·Baley A FongDavid W Wood

❮ Previous
Next ❯

Citations

Jan 27, 2011·Biotechnology Letters·Yifeng Li
Apr 23, 2021·Journal of Agricultural and Food Chemistry·Yifeng ZhangYingfeng An

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.