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Article: High level expression, purification and characterization of active fusion human C1q and tumor necrosis factor related protein 2 (hCTRP2) in Escherichia coli
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TitleHigh level expression, purification and characterization of active fusion human C1q and tumor necrosis factor related protein 2 (hCTRP2) in Escherichia coli
 
AuthorsLi, H3
Gao, X3
Zhou, Y3
Li, N3
Ge, C3
Hui, X1
Wang, Y1 3
Xu, A1 3
Jin, S2
Wu, D3
 
KeywordsEscherichia coli
Expression and purification
Fusion protein
Human C1q and tumor necrosis factor related protein 2 (hCTRP2)
Pichia pastoris
 
Issue Date2011
 
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/yprep
 
CitationProtein Expression And Purification, 2011, v. 79 n. 1, p. 1-6 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.pep.2011.03.013
 
AbstractC1q and tumor necrosis factor related proteins (CTRPs) are a family of adiponectin paralogues. Among them, CTRP2 is the only CTRP protein that has been shown to possess similar biological activities as adiponectin. To further explore the physiological roles of human CTRP2 and its mechanisms of action, hCTRP2 gene was expressed in Escherichia coli and Pichia pastoris, respectively. In the P. pastoris expression system, recombinant hCTRP2 could be secreted into the culture medium under induction condition, however, the resultant recombinant protein was highly unstable, resulting two main degradation products with molecular masses of approximately 20 and 26 kDa, respectively. In the E. coli expression system, a large amount of soluble thioredoxin (Trx)-hCTRP2 fusion protein could be produced, which accounts about 42% of the total soluble bacterial proteins. The recombinant Trx-hCTRP2 fusion protein was purified to an approximately 95% purity using Ni-NTA affinity chromatography and Superdex G-75 column with a yield of about 15 mg/l protein from 1 l bacterial culture. The purified recombinant Trx-hCTRP2 was shown to be active under in vitro assay conditions. © 2011 Elsevier Inc. All rights reserved.
 
ISSN1046-5928
2013 Impact Factor: 1.508
 
DOIhttp://dx.doi.org/10.1016/j.pep.2011.03.013
 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorLi, H
 
dc.contributor.authorGao, X
 
dc.contributor.authorZhou, Y
 
dc.contributor.authorLi, N
 
dc.contributor.authorGe, C
 
dc.contributor.authorHui, X
 
dc.contributor.authorWang, Y
 
dc.contributor.authorXu, A
 
dc.contributor.authorJin, S
 
dc.contributor.authorWu, D
 
dc.date.accessioned2012-09-05T05:30:46Z
 
dc.date.available2012-09-05T05:30:46Z
 
dc.date.issued2011
 
dc.description.abstractC1q and tumor necrosis factor related proteins (CTRPs) are a family of adiponectin paralogues. Among them, CTRP2 is the only CTRP protein that has been shown to possess similar biological activities as adiponectin. To further explore the physiological roles of human CTRP2 and its mechanisms of action, hCTRP2 gene was expressed in Escherichia coli and Pichia pastoris, respectively. In the P. pastoris expression system, recombinant hCTRP2 could be secreted into the culture medium under induction condition, however, the resultant recombinant protein was highly unstable, resulting two main degradation products with molecular masses of approximately 20 and 26 kDa, respectively. In the E. coli expression system, a large amount of soluble thioredoxin (Trx)-hCTRP2 fusion protein could be produced, which accounts about 42% of the total soluble bacterial proteins. The recombinant Trx-hCTRP2 fusion protein was purified to an approximately 95% purity using Ni-NTA affinity chromatography and Superdex G-75 column with a yield of about 15 mg/l protein from 1 l bacterial culture. The purified recombinant Trx-hCTRP2 was shown to be active under in vitro assay conditions. © 2011 Elsevier Inc. All rights reserved.
 
dc.description.naturelink_to_subscribed_fulltext
 
dc.identifier.citationProtein Expression And Purification, 2011, v. 79 n. 1, p. 1-6 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.pep.2011.03.013
 
dc.identifier.citeulike9126065
 
dc.identifier.doihttp://dx.doi.org/10.1016/j.pep.2011.03.013
 
dc.identifier.epage6
 
dc.identifier.hkuros206391
 
dc.identifier.issn1046-5928
2013 Impact Factor: 1.508
 
dc.identifier.issue1
 
dc.identifier.pmid21453774
 
dc.identifier.scopuseid_2-s2.0-79960133528
 
dc.identifier.spage1
 
dc.identifier.urihttp://hdl.handle.net/10722/163385
 
dc.identifier.volume79
 
dc.languageeng
 
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/yprep
 
dc.publisher.placeUnited States
 
dc.relation.ispartofProtein Expression and Purification
 
dc.relation.referencesReferences in Scopus
 
dc.subject.meshCloning, Molecular
 
dc.subject.meshEscherichia Coli - Genetics
 
dc.subject.meshGene Expression
 
dc.subject.meshHumans
 
dc.subject.meshPichia - Genetics
 
dc.subject.meshProteins - Genetics - Isolation & Purification - Metabolism
 
dc.subject.meshRecombinant Fusion Proteins - Genetics - Isolation & Purification - Metabolism
 
dc.subjectEscherichia coli
 
dc.subjectExpression and purification
 
dc.subjectFusion protein
 
dc.subjectHuman C1q and tumor necrosis factor related protein 2 (hCTRP2)
 
dc.subjectPichia pastoris
 
dc.titleHigh level expression, purification and characterization of active fusion human C1q and tumor necrosis factor related protein 2 (hCTRP2) in Escherichia coli
 
dc.typeArticle
 
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<description.abstract>C1q and tumor necrosis factor related proteins (CTRPs) are a family of adiponectin paralogues. Among them, CTRP2 is the only CTRP protein that has been shown to possess similar biological activities as adiponectin. To further explore the physiological roles of human CTRP2 and its mechanisms of action, hCTRP2 gene was expressed in Escherichia coli and Pichia pastoris, respectively. In the P. pastoris expression system, recombinant hCTRP2 could be secreted into the culture medium under induction condition, however, the resultant recombinant protein was highly unstable, resulting two main degradation products with molecular masses of approximately 20 and 26 kDa, respectively. In the E. coli expression system, a large amount of soluble thioredoxin (Trx)-hCTRP2 fusion protein could be produced, which accounts about 42% of the total soluble bacterial proteins. The recombinant Trx-hCTRP2 fusion protein was purified to an approximately 95% purity using Ni-NTA affinity chromatography and Superdex G-75 column with a yield of about 15 mg/l protein from 1 l bacterial culture. The purified recombinant Trx-hCTRP2 was shown to be active under in vitro assay conditions. &#169; 2011 Elsevier Inc. All rights reserved.</description.abstract>
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Author Affiliations
  1. The University of Hong Kong
  2. University of Florida
  3. Chinese Academy of Sciences