About: pC6-2/caspase-6 system to purify glutathione-S-transferase-free recombinant fusion proteins expressed in Escherichia coli

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About: pC6-2/caspase-6 system to purify glutathione-S-transferase-free recombinant fusion proteins expressed in Escherichia coli Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	pC6-2/caspase-6 system to purify glutathione-S-transferase-free recombinant fusion proteins expressed in Escherichia coli
Creator	Galande, Sanjeev Patole, Milind Jayakumar, Cyril Purbey, Kumar
Source	Medline; PMC
abstract	Glutathione-S-transferase (GST) fusion protein expression vectors are often employed for the expression and purification of proteins in Escherichia coli. GST is then removed by site-specific proteolysis using thrombin. However, the presence of internal thrombin cleavage sites in expressed proteins can severely affect the purification of intact proteins. Cysteine-dependent aspartate-specific proteases (caspases) are efficient enzymes with defined substrate specificity. Unlike most of the proteases used for the removal of affinity tags, caspases do not leave any amino acids at the amino-terminus of cleaved proteins. We have engineered the caspase-6 site VEMD in a pGEX vector to give the pC6-2 vector. The caspase-6 can be easily removed after cleavage. Here, we describe the detailed protocol for purifying proteins using our pC6-2/caspase-6 expression and purification system. The cleavage by caspase-6 occurs in <30 min and the entire procedure can be completed in 2 d.
has issue date	2006-11-16(xsd:dateTime)
bibo:doi	10.1038/nprot.2006.310
bibo:pmid	17487164
has license	no-cc
sha1sum (hex)	3ad9af7b8efd7b1ad327af69ef21841a596c9477
schema:url	https://doi.org/10.1038/nprot.2006.310
resource representing a document's title	pC6-2/caspase-6 system to purify glutathione-S-transferase-free recombinant fusion proteins expressed in Escherichia coli
has PubMed Central identifier	PMC7086577
has PubMed identifier	17487164
schema:publication	Nat Protoc
resource representing a document's body	covid:3ad9af7b8efd7b1ad327af69ef21841a596c9477#body_text
is schema:about of	named entity 'ESCHERICHIA COLI' named entity 'cleavage' named entity 'proteins' named entity 'proteins' named entity 'proteases' named entity 'purifying' named entity 'PURIFY' named entity 'GLUTATHIONE' named entity 'SITE' named entity 'PROTOCOL' named entity 'PURIFICATION' named entity 'EMPLOYED' named entity 'PROTEOLYSIS' named entity 'ENGINEERED' named entity 'EFFICIENT' named entity 'FREE' named entity 'ENTIRE' named entity 'CASPASE-6' named entity 'USED' named entity 'TRANSFERASE' named entity 'INTACT' named entity 'CLEAVED' named entity 'SPECIFIC' named entity 'EXPRESSION' named entity 'DEFINED' named entity 'REMOVAL' named entity 'PROTEINS' named entity 'RECOMBINANT FUSION PROTEINS' named entity 'AMINO ACIDS' named entity 'EXPRESSED' named entity 'HAVE' named entity 'DO NOT' named entity 'FUSION PROTEIN EXPRESSION' named entity 'ESCHERICHIA COLI' named entity 'ASPARTATE' named entity 'ENZYMES' named entity 'GST' named entity 'PROTEASES' named entity 'PROCEDURE' named entity 'CLEAVAGE' named entity 'DESCRIBE' named entity 'USING' named entity 'CYSTEINE' named entity 'EASILY' named entity 'SYSTEM' named entity 'SEVERELY' named entity 'CASPASES' named entity 'EXPRESSED' named entity 'O30' named entity 'DEPENDENT' named entity 'SUBSTRATE SPECIFICITY' named entity 'TRANSFERASE' named entity 'TAGS' named entity 'PRESENCE OF' named entity 'OUR' named entity 'MIN' named entity 'PURIFYING' named entity 'MOST OF' named entity 'DETAILED' named entity 'AMINO-TERMINUS' named entity 'GLUTATHIONE' named entity 'AFFINITY' named entity 'THROMBIN' named entity 'LEAVE' named entity 'REMOVED' named entity 'AFFECT' named entity 'INTERNAL' named entity 'VECTOR' named entity 'PC6' named entity 'COMPLETED' named entity 'EXPRESSION VECTORS' named entity 'PC6' named entity 'HERE' named entity 'SYSTEM' covid:arg/3ad9af7b8efd7b1ad327af69ef21841a596c9477 named entity 'expression' named entity 'affinity tags' named entity 'purification'

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