Article: High level expression, purification and characterization of active fusion human C1q and tumor necrosis factor related protein 2 (hCTRP2) in Escherichia coli

File Download Links for fulltext
(May Require Subscription)
Supplementary
  • Basic View
  • Metadata View
  • XML View
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
2011 Impact Factor: 1.587
2011 SCImago Journal Rankings: 0.179
DOIhttp://dx.doi.org/10.1016/j.pep.2011.03.013
ReferencesReferences in Scopus
DC Field
Value
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
2011 Impact Factor: 1.587
2011 SCImago Journal Rankings: 0.179
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
Author Affiliations
  1. The University of Hong Kong
  2. University of Florida
  3. Chinese Academy of Sciences