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Conference Paper: Zebrafish telomerase catalytic subunit: Molecular cloning and characterization of a ribonucleoprotein reverse transcriptase gene highly expressed in the retina

TitleZebrafish telomerase catalytic subunit: Molecular cloning and characterization of a ribonucleoprotein reverse transcriptase gene highly expressed in the retina
Authors
Issue Date2005
PublisherAssociation for Research in Vision and Ophthalmology
Citation
The Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO), Fort Lauderdale, Florida, 1-4 May 2005. In Investigative Ophthalmology & Visual Science, 2005, v. 46 n. 13, p. 5329 How to Cite?
AbstractAbstract: : Purpose: To clone and sequence the zebrafish TERT gene and to study the expression pattern of TERT and telomerase activity in zebrafish retina. Methods: Primers identical to those used originally to amplify and clone the mouse TERT sequence were retrieved from tBLASTn, and were used to amplify zebrafish genomic DNA by PCR. The amplified product was then cloned into the pDNR–CMV vector using the In–Fusion PCR cloning kit (BD Bioscience). All sequencing analyses were carried out at the Genome Research Centre, at the University of Hong Kong. Western blotting, immunohistochemistry, RT–PCR and telomeric repeats amplification protocol (TRAP) assay are performed to study TERT mRNA and protein expression and telomerase activity in zebrafish retina. Results: Based on the amino acid sequence of mouse TERT, a full–length telomerase reverse transcriptase cDNA of zebrafish has been isolated and cloned. The deduced protein sequence contains 1091 amino–acid residues and a predicted molecular mass of 126 kDa. Multiple alignments reveal that the protein sequence bears the conserved motifs and residues found in TERT of other species. RT–PCR and TRAP assay detect TERT mRNA expression and telomerase activity in all tissues examined, including retina and brain. Presence of telomerase activity indicates that a fully functional form of telomerase can be found in retina. Through immunohistochemistry, the protein was localized to cells in the ganglion cell layer, and to inner and outer nuclear layers of adult zebrafish retina. Conclusions: The presence of telomerase in most somatic tissues in zebrafish is consistent with the high proliferation capacity of cells in organs of teleost fish which grow throughout their life with little senescence. However, retinal neurons in adult teleost are post–mitotic cells and do not divide under normal situation. The expression of TERT and telomerase activity in the retina suggests that telomerase has functions other than elongation of telomere. Functional studies can now be carried out to shed further light on the role of TERT in vision.
Persistent Identifierhttp://hdl.handle.net/10722/95229
ISSN

 

DC FieldValueLanguage
dc.contributor.authorYip, HKFen_HK
dc.contributor.authorLau, WMen_HK
dc.contributor.authorTsao, GSWen_HK
dc.contributor.authorSo, KFen_HK
dc.contributor.authorWong, AOLen_HK
dc.date.accessioned2010-09-25T15:55:41Z-
dc.date.available2010-09-25T15:55:41Z-
dc.date.issued2005en_HK
dc.identifier.citationThe Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO), Fort Lauderdale, Florida, 1-4 May 2005. In Investigative Ophthalmology & Visual Science, 2005, v. 46 n. 13, p. 5329-
dc.identifier.issn1552-5783-
dc.identifier.urihttp://hdl.handle.net/10722/95229-
dc.description.abstractAbstract: : Purpose: To clone and sequence the zebrafish TERT gene and to study the expression pattern of TERT and telomerase activity in zebrafish retina. Methods: Primers identical to those used originally to amplify and clone the mouse TERT sequence were retrieved from tBLASTn, and were used to amplify zebrafish genomic DNA by PCR. The amplified product was then cloned into the pDNR–CMV vector using the In–Fusion PCR cloning kit (BD Bioscience). All sequencing analyses were carried out at the Genome Research Centre, at the University of Hong Kong. Western blotting, immunohistochemistry, RT–PCR and telomeric repeats amplification protocol (TRAP) assay are performed to study TERT mRNA and protein expression and telomerase activity in zebrafish retina. Results: Based on the amino acid sequence of mouse TERT, a full–length telomerase reverse transcriptase cDNA of zebrafish has been isolated and cloned. The deduced protein sequence contains 1091 amino–acid residues and a predicted molecular mass of 126 kDa. Multiple alignments reveal that the protein sequence bears the conserved motifs and residues found in TERT of other species. RT–PCR and TRAP assay detect TERT mRNA expression and telomerase activity in all tissues examined, including retina and brain. Presence of telomerase activity indicates that a fully functional form of telomerase can be found in retina. Through immunohistochemistry, the protein was localized to cells in the ganglion cell layer, and to inner and outer nuclear layers of adult zebrafish retina. Conclusions: The presence of telomerase in most somatic tissues in zebrafish is consistent with the high proliferation capacity of cells in organs of teleost fish which grow throughout their life with little senescence. However, retinal neurons in adult teleost are post–mitotic cells and do not divide under normal situation. The expression of TERT and telomerase activity in the retina suggests that telomerase has functions other than elongation of telomere. Functional studies can now be carried out to shed further light on the role of TERT in vision.-
dc.languageengen_HK
dc.publisherAssociation for Research in Vision and Ophthalmology-
dc.relation.ispartofInvestigative Ophthalmology & Visual Scienceen_HK
dc.titleZebrafish telomerase catalytic subunit: Molecular cloning and characterization of a ribonucleoprotein reverse transcriptase gene highly expressed in the retinaen_HK
dc.typeConference_Paperen_HK
dc.identifier.emailYip, HKF: hkfyip@hku.hken_HK
dc.identifier.emailTsao, GSW: gswtsao@hkucc.hku.hken_HK
dc.identifier.emailSo, KF: hrmaskf@hkucc.hku.hken_HK
dc.identifier.emailWong, AOL: olwong@HKUCC.hku.hken_HK
dc.identifier.authorityTsao, GSW=rp00399en_HK
dc.identifier.authoritySo, KF=rp00329en_HK
dc.identifier.authorityWong, AOL=rp00806en_HK
dc.identifier.hkuros107492en_HK

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