PMID: 20636269Jul 20, 2010Paper

Purification of core enzyme of Escherichia coli RNA polymerase by affinity chromatography

Biochemistry. Biokhimii︠a︡
Yu A KhodakV L Drutsa

Abstract

A method for isolation of a highly purified preparation of E. coli RNA polymerase core enzyme was developed based on IMPACT technology and dissociation of the RNA polymerase complex with sigma(70) subunit. Washing of the immobilized RNA polymerase with 5-10 mM solution of glutamate (pH 5.0-5.5) completely removed the sigma(70) subunit from the holoenzyme and decreased amounts of protein admixtures. The possibility of reconstruction of the RNA polymerase holoenzyme directly on the affinity column was demonstrated. Activities of the resulting RNAP core enzyme preparations were tested by in vitro transcription. Some amino acids and their mixtures were shown to influence the in vitro transcription. The findings indicate that changes in the transcription efficiency in the presence of amino acids should be associated with a specific destruction of the interaction between sigma(70) subunit and the core enzyme.

References

May 23, 1995·Proceedings of the National Academy of Sciences of the United States of America·H TangR H Ebright
Jan 9, 1996·Proceedings of the National Academy of Sciences of the United States of America·D P GreinerC F Meares
Jan 1, 1996·Methods in Enzymology·N Fujita, A Ishihama
Apr 17, 1998·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·H Nagai, N Shimamoto
May 30, 1998·Proceedings of the National Academy of Sciences of the United States of America·J T OwensC F Meares
Nov 18, 1998·FEMS Microbiology Reviews·M M Wösten
Oct 6, 2000·Annual Review of Microbiology·A Ishihama
Apr 3, 2001·Current Opinion in Microbiology·R R Burgess, L Anthony
Feb 13, 2003·Current Opinion in Structural Biology·Katsuhiko S Murakami, Seth A Darst
May 7, 2003·Current Opinion in Microbiology·Sergei Borukhov, Evgeny Nudler
Oct 7, 2004·Journal of Molecular Biology·Kati GeszvainRobert Landick
Mar 12, 2005·Proceedings of the National Academy of Sciences of the United States of America·Bryce E NickelsAnn Hochschild
Nov 16, 2005·Molecular Cell·Rachel Anne MooneyRobert Landick
Apr 12, 2006·Genome Research·Mohammad ArifuzzamanHirotada Mori
Dec 5, 2008·Biochemistry·Jay D Gralla, Yi-Xin Huo

❮ Previous
Next ❯

Citations

Sep 6, 2011·Nucleosides, Nucleotides & Nucleic Acids·Olga Nikolaevna KorolevaValeriy Lvovich Drutsa
Jun 28, 2011·Nanomedicine : Nanotechnology, Biology, and Medicine·Evgeniy V DubrovinValeriy L Drutsa

❮ 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.