scholarly journals Improved two‐stage protein expression and purification via autoinduction of both autolysis and auto DNA/RNA hydrolysis conferred by phage lysozyme and DNA/RNA endonuclease

2020 ◽  
Vol 117 (9) ◽  
pp. 2852-2860 ◽  
Author(s):  
Romel Menacho‐Melgar ◽  
Eirik A. Moreb ◽  
John P. Efromson ◽  
Tian Yang ◽  
Jennifer N. Hennigan ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression And Purification ◽  
Two Stage ◽  
Protein Expression And Purification ◽  
Rna Hydrolysis ◽  
Rna Endonuclease

scholarly journals Improved, two-stage protein expression and purification via autoinduction of both autolysis and auto DNA/RNA hydrolysis conferred by phage lysozyme and DNA/RNA endonuclease

2020 ◽  
Author(s):  
Romel Menacho-Melgar ◽  
Eirik A. Moreb ◽  
John P. Efromson ◽  
Michael D. Lynch
Keyword(s):  
Protein Expression ◽  
Membrane Integrity ◽  
Heterologous Protein Expression ◽  
List Type ◽  
Two Stage ◽  
Microtiter Plates ◽  
Thaw Cycle ◽  
Protein Expression And Purification ◽  
Phosphate Depletion ◽  
Rna Endonuclease

AbstractWe report improved release of recombinant proteins in E. coli, which relies on combined cellular autolysis and DNA/RNA autohydrolysis, conferred by the tightly controlled autoinduction of both phage lysozyme and the non specific DNA/RNA endonuclease from S. marcescens. Autoinduction occurs in a two-stage process wherein heterologous protein expression and autolysis enzymes are induced upon entry into stationary phase by phosphate depletion. Cytoplasmic lysozyme and periplasmic endonuclease are kept from inducing lysis until membrane integrity is disrupted. Post cell harvest, the addition of detergent (0.1% Triton-X100) and a single 30 minutes freezer thaw cycle results in > 90% release of protein (GFP). This cellular lysis is accompanied by complete oligonucleotide hydrolysis. The approach has been validated for shake flask cultures, high throughput cultivation in microtiter plates and larger scale stirred-tank bioreactors. This tightly controlled system enables robust growth and resistance to lysis in routine media when cells are propagated and autolysis/hydrolysis genes are only induced upon phosphate depletion.HighlightsAutoinduction of both cell lysis and nucleotide hydrolysis>90 % lysis and DNA degradationStrains are stable to lysis in the absence of phosphate depletion.

pA-Hia5 Protein Expression and Purification v1

protocols.io ◽  
2021 ◽  
Author(s):  
Nicolas Altemose ◽  
Annie Maslan ◽  
Owen Smith ◽  
Kousik Sundararajan ◽  
Rachel Brown ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression And Purification ◽  
Protein Expression And Purification

scholarly journals A baculovirus-mediated strategy for full-length plant virus coat protein expression and purification

2013 ◽  
Vol 10 (1) ◽  
pp. 262 ◽  
Author(s):  
Daniel Mendes Ardisson-Araújo ◽  
Juliana Rocha ◽  
Márcio Hedil da Costa ◽  
Anamélia Bocca ◽  
André Dusi ◽  
...  
Keyword(s):  
Coat Protein ◽  
Protein Expression ◽  
Plant Virus ◽  
Full Length ◽  
Expression And Purification ◽  
Virus Coat Protein ◽  
Protein Expression And Purification ◽  
Virus Coat

scholarly journals Peptides from the SARS-associated coronavirus as tags for protein expression and purification

2008 ◽  
Vol 61 (2) ◽  
pp. 138-141 ◽  
Author(s):  
Steffi Gruschke ◽  
Bianca M. Bußmann ◽  
Sven Reiche ◽  
Christian Jassoy
Keyword(s):  
Protein Expression ◽  
Expression And Purification ◽  
Protein Expression And Purification
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