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Production and characterisation of gilthead sea bream (Sparus auratus) recombinant parathyroid hormone related protein

2005Journal article Restricted access
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dc.contributor.authorAnjos, Liliana
dc.contributor.authorRotllant, J.
dc.contributor.authorGuerreiro, P. M.
dc.contributor.authorHang, X. M.
dc.contributor.authorCanario, Adelino V. M.
dc.contributor.authorBalment, R.
dc.contributor.authorPower, Deborah
dc.date.accessioned2014-05-17T09:13:47Z
dc.date.available2014-05-17T09:13:47Z
dc.date.issued2005
dc.date.updated2014-05-13T10:05:43Z
dc.description.abstractThe production and puriWcation of gilthead sea bream recombinant parathyroid hormone related protein [sbPTHrP(1–125)] using an Escherichia coli system and one step puriWcation process with continuous elution gel electrophoresis is reported. The cDNA encoding sbPTHrP(1–125) was cloned into a prokaryotic expression vector pET-11a. The recombinant plasmid was used to transfect E. coli BL21(DE3) pLysS and sbPTHrP(1–125) synthesis was induced by addition of 1mM isopropyl- -D-thiogalactopyranoside. The rapid one step isolation method gave pure sbPTHrP(1–125) as judged by SDS–PAGE and yielded up to 40mg/L of culture medium (3.3mg protein/g of bacteria). The bioactivity of recombinant sbPTHrP(1–125) assessed using an in vitro scale bioassay was found to be equipotent to PTHrP(1–34) in stimulating cAMP accumulation. Assessment of the immunological reactivity of the isolated protein by Western blot revealed it cross-reacts with antisera speciWc for the N-terminal and C-terminal region of PTHrP. In a radioimmunoassay speciWc for piscine N-terminal (1–34 aa) PTHrP, the recombinant sbPTHrP(1–125) was equipotent with PTHrP(1–34) in displacing labelled 125I-PTHrP(1–36) PTHrP from the antisera. The availability of recombinant sbPTHrP will allow the development of region speciWc assays and studies aimed at deWning post-secretory processing of this protein and its biological activity in fish.por
dc.identifier.citationAnjos, L.; Rotlant, J.; Guerreiro, P. M.; Hang, X. M.; Canario, A. V. M.; Balment, R.; Power, D. M.Production and characterisation of gilthead sea bream (Sparus auratus) recombinant parathyroid hormone related protein, General and Comparative Endocrinology, 143, 1, 57-65, 2005.por
dc.identifier.doihttp://dx.doi.org/10.1016/j.ygcen.2005.02.020
dc.identifier.issn0016-6480
dc.identifier.otherAUT: ACA00258; DPO00386;
dc.identifier.urihttp://hdl.handle.net/10400.1/3989
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherElsevierpor
dc.relation.publisherversionhttp://www.sciencedirect.com/science/article/pii/S0016648005000651por
dc.subjectRecombinant parathyroid hormone related proteinpor
dc.subjectE. coli productionpor
dc.subjectBiological activitypor
dc.subjectImmunological activitypor
dc.subjectTeleostpor
dc.titleProduction and characterisation of gilthead sea bream (Sparus auratus) recombinant parathyroid hormone related proteinpor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage65por
oaire.citation.issue1
oaire.citation.startPage57por
oaire.citation.titleGeneral and Comparative Endocrinologypor
oaire.citation.volume143por
person.familyNameAnjos Guerreiro
person.familyNameGuerreiro da Costa Guerreiro
person.familyNameCanario
person.familyNamePower
person.givenNameLiliana Isabel Tomé
person.givenNamePedro Miguel
person.givenNameAdelino
person.givenNameDeborah Mary
person.identifierAAK-3800-2020
person.identifierA-2539-2009
person.identifier143624
person.identifier.ciencia-id6D18-2D1C-A45C
person.identifier.ciencia-id5C13-965D-3148
person.identifier.ciencia-id1F1E-D3B3-F804
person.identifier.ciencia-id891A-8A44-3CAE
person.identifier.orcid0000-0001-9474-522X
person.identifier.orcid0000-0001-5371-7919
person.identifier.orcid0000-0002-6244-6468
person.identifier.orcid0000-0003-1366-0246
person.identifier.ridC-7942-2009
person.identifier.scopus-author-id8575757700
person.identifier.scopus-author-id56568523700
person.identifier.scopus-author-id7101806760
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor
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