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Article: Molecular cloning and expression studies of a prolactin receptor in goldfish (Carassius auratus)

TitleMolecular cloning and expression studies of a prolactin receptor in goldfish (Carassius auratus)
Authors
KeywordscDNA cloning
Functional expression
Goldfish
Prolactin receptor
Tissue distribution
Issue Date2000
PublisherElsevier Inc. The Journal's web site is located at http://www.elsevier.com/locate/lifescie
Citation
Life Sciences, 2000, v. 66 n. 7, p. 593-605 How to Cite?
AbstractA full-length cDNA clone, of a size of 4.6 kb, for the goldfish prolactin receptor has been isolated. This cDNA clone encodes a protein of 600 amino acids homologous to prolactin receptors of other species. A Kyte- Doolittle hydropathy analysis of the receptor indicates that the translated protein consists of a signal peptide of 22 amino acids, an extracellular domain of 228 amino acids, a single transmembrane domain of 24 amino acids, and an intracellular domain of 346 amino acids. Several characteristic landmarks of prolactin receptor could be identified in this clone. These include the four conserved cysteine residues and the WS motif within the extracellular domain, and the box 1 and box 2 regions of the intracellular domain. Among all the prolactin receptor sequences known to date, this clone bears the closest resemblance to the tilapia prolactin receptor, although homology between these two fish prolactin receptors is rather low. There are only 57.4% of nucleotide and 48.3% of amino acid sequence identities between these two fish receptors. This receptor cDNA was transfected into CHO-K1 cells for functional analysis. RT-PCR analysis with a pair of gene specific primers indicate that the receptor was transcribed in the transfected cells. Using a cell proliferation assay based on the reduction of the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, the receptor transfected CHO-K1 cells can be stimulated to proliferate upon the addition of ovine prolactin in the culture medium. The tissue distribution of the prolactin receptor in goldfish was studied by RT-PCR/Southern analysis and by Northern analysis. The results indicated that the receptor is expressed mostly in the kidney, the gill and the intestine of goldfish, corroborating with the osmoregulatory role of prolactin in fish. In addition, an appreciable level of the receptor is also found in the brain and gonads of goldfish. Northern analysis showed that there are two transcript sizes, a major 4.6 kb and a minor 3.5 kb mRNAs, in the kidney, gill and intestine.
Persistent Identifierhttp://hdl.handle.net/10722/84687
ISSN
2015 Impact Factor: 2.685
2015 SCImago Journal Rankings: 1.056
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorTse, DLYen_HK
dc.contributor.authorChow, BKCen_HK
dc.contributor.authorChan, CBen_HK
dc.contributor.authorLee, LTOen_HK
dc.contributor.authorCheng, CHKen_HK
dc.date.accessioned2010-09-06T08:55:57Z-
dc.date.available2010-09-06T08:55:57Z-
dc.date.issued2000en_HK
dc.identifier.citationLife Sciences, 2000, v. 66 n. 7, p. 593-605en_HK
dc.identifier.issn0024-3205en_HK
dc.identifier.urihttp://hdl.handle.net/10722/84687-
dc.description.abstractA full-length cDNA clone, of a size of 4.6 kb, for the goldfish prolactin receptor has been isolated. This cDNA clone encodes a protein of 600 amino acids homologous to prolactin receptors of other species. A Kyte- Doolittle hydropathy analysis of the receptor indicates that the translated protein consists of a signal peptide of 22 amino acids, an extracellular domain of 228 amino acids, a single transmembrane domain of 24 amino acids, and an intracellular domain of 346 amino acids. Several characteristic landmarks of prolactin receptor could be identified in this clone. These include the four conserved cysteine residues and the WS motif within the extracellular domain, and the box 1 and box 2 regions of the intracellular domain. Among all the prolactin receptor sequences known to date, this clone bears the closest resemblance to the tilapia prolactin receptor, although homology between these two fish prolactin receptors is rather low. There are only 57.4% of nucleotide and 48.3% of amino acid sequence identities between these two fish receptors. This receptor cDNA was transfected into CHO-K1 cells for functional analysis. RT-PCR analysis with a pair of gene specific primers indicate that the receptor was transcribed in the transfected cells. Using a cell proliferation assay based on the reduction of the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, the receptor transfected CHO-K1 cells can be stimulated to proliferate upon the addition of ovine prolactin in the culture medium. The tissue distribution of the prolactin receptor in goldfish was studied by RT-PCR/Southern analysis and by Northern analysis. The results indicated that the receptor is expressed mostly in the kidney, the gill and the intestine of goldfish, corroborating with the osmoregulatory role of prolactin in fish. In addition, an appreciable level of the receptor is also found in the brain and gonads of goldfish. Northern analysis showed that there are two transcript sizes, a major 4.6 kb and a minor 3.5 kb mRNAs, in the kidney, gill and intestine.en_HK
dc.languageengen_HK
dc.publisherElsevier Inc. The Journal's web site is located at http://www.elsevier.com/locate/lifescieen_HK
dc.relation.ispartofLife Sciencesen_HK
dc.rightsLife Sciences. Copyright © Elsevier Inc.en_HK
dc.subjectcDNA cloningen_HK
dc.subjectFunctional expressionen_HK
dc.subjectGoldfishen_HK
dc.subjectProlactin receptoren_HK
dc.subjectTissue distributionen_HK
dc.titleMolecular cloning and expression studies of a prolactin receptor in goldfish (Carassius auratus)en_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0024-3205&volume=66&spage=593&epage=605&date=2000&atitle=Molecular+Cloning+and+Expression+Studies+of+A+Prolactin+Receptor+in+Goldfish+(Carassius+auratus)en_HK
dc.identifier.emailChow, BKC: bkcc@hku.hken_HK
dc.identifier.emailLee, LTO: ltolee2@hkucc.hku.hken_HK
dc.identifier.authorityChow, BKC=rp00681en_HK
dc.identifier.authorityLee, LTO=rp00727en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/S0024-3205(99)00632-3en_HK
dc.identifier.pmid10794515en_HK
dc.identifier.scopuseid_2-s2.0-0034614450en_HK
dc.identifier.hkuros47973en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0034614450&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume66en_HK
dc.identifier.issue7en_HK
dc.identifier.spage593en_HK
dc.identifier.epage605en_HK
dc.identifier.isiWOS:000084742200005-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridTse, DLY=7101916515en_HK
dc.identifier.scopusauthoridChow, BKC=7102826193en_HK
dc.identifier.scopusauthoridChan, CB=7404813651en_HK
dc.identifier.scopusauthoridLee, LTO=8367269000en_HK
dc.identifier.scopusauthoridCheng, CHK=7404798014en_HK

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