Abstract：
:PagP, a beta-barrel membrane protein found in Gram-negative bacteria, expresses robustly in inclusion bodies when its signal sequence is removed. We have developed a new fusion protein expression system based on PagP and demonstrated its utility in the expression of the unstructured N-terminal region of human cardiac troponin I (residues 1-71). A yield of 100mg fusion protein per liter M9 minimal media was obtained. The troponin I fragment was removed from PagP using cyanogen bromide cleavage at methionine residues followed by nickel affinity chromatography. We further demonstrate that optimal cleavage requires complete reduction of methionine residues prior to cyanogen bromide treatment, and this is effectively accomplished using potassium iodide under acidic conditions. The PagP-based fusion protein system is more effective at targeting proteins into inclusion bodies than a commercially available system that uses ketosteroid isomerase; it thus represents an important advance for producing large quantities of unfolded peptides or proteins in Escherichia coli.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Hwang PM,Pan JS,Sykes BDdoi
10.1016/j.pep.2012.07.007subject
Has Abstractpub_date
2012-09-01 00:00:00pages
148-51issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(12)00203-3journal_volume
85pub_type
杂志文章abstract：:Rab GTPases are post-translationally geranylgeranylated on their C-terminal cysteines by Rab geranylgeranyl transferase (RabGGTase) and this modification is essential for their biological activity. Rab Escort Protein (REP) is a molecular chaperone that assists in the prenylation reaction carried out by RabGGTase. Muta...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00506-5
更新日期：2002-10-01 00:00:00
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2010.12.008
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.03.021
更新日期：2005-07-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1996.0705
更新日期：1997-06-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2018.08.010
更新日期：2019-01-01 00:00:00
abstract：UNLABELLED:Recombinant protein production is a significant biotechnological process as it allows researchers to produce a specific protein in desired quantities. Escherichia coli (E. coli) is the most popular heterologous expression host for the production of recombinant proteins due to its advantages such as low cost,...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2013.11.014
更新日期：2014-03-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.10.008
更新日期：2007-03-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.01.019
更新日期：2004-06-01 00:00:00
abstract：:Proteins are essential throughout the biological and biomedical sciences and the purification strategies of proteins of interest have advanced over centuries. Elastin-like polypeptides (ELPs) are compound polymers that have recently been highlighted for their sharp and reversible phase transition property when heated ...
journal_title：Protein expression and purification
pub_type： 杂志文章,评审
doi：10.1016/j.pep.2018.09.006
更新日期：2019-01-01 00:00:00
abstract：:Transcription factor HIF-1 is a key regulator in cellular adaptation to hypoxia. HIF prolyl hydroxylases (PHDs) control HIF-1 accumulation by hydroxylation dependent on molecular oxygen. Due to this regulation, PHDs have been pointed out as potential drug targets. We have purified catalytically active human PHD3 after...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.02.018
更新日期：2007-07-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2011.06.012
更新日期：2011-11-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2001.1586
更新日期：2002-04-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
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更新日期：2015-11-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.06.011
更新日期：2006-11-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1999.1158
更新日期：1999-12-01 00:00:00
abstract：:A rabbit liver cDNA library in phage lambdagt10 was screened using the coding cDNA for human cytosolic serine hydroxymethyltransferase. A clone of 1754 bp was isolated and the nucleotide sequence showed an open reading frame of 1455 bp, which coded for rabbit cytosolic serine hydroxymethyltransferase and was flanked b...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1998.0890
更新日期：1998-07-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1996.0113
更新日期：1996-11-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2001.1553
更新日期：2002-02-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.01.012
更新日期：2014-04-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.01.022
更新日期：2006-08-01 00:00:00
abstract：:The role of the termini of protein sequences is often perturbed by remnant amino acids after the specific protease cleavage of the affinity tags and/or by the amino acids encoded by the plasmid at/around the restriction enzyme sites used to insert the genes. Here we describe a method for affinity purification of a met...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.03.009
更新日期：2019-07-01 00:00:00
abstract：:Developmentally regulated G-proteins (DRGs) are a highly conserved family of GTP-binding proteins found in archaea, plants, fungi and animals, indicating important roles in fundamental pathways. Their function is poorly understood, but they have been implicated in cell division, proliferation, and growth, as well as s...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.05.009
更新日期：2009-10-01 00:00:00
