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Molecular analysis of T-DNA insertion mutants identified putative regulatory elements in the AtTERT gene

Analysis of plants bearing a T-DNA insertion is a potent tool of modern molecular biology, providing valuable information about the function and involvement of genes in metabolic pathways. A collection of 12 Arabidopsis thaliana lines with T-DNA insertions in the gene coding for the catalytic subuni...

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Autores principales: Fojtová, Miloslava, Peška, Vratislav, Dobšáková, Zuzana, Mozgová, Iva, Fajkus, Jiří, Sýkorová, Eva
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3223050/
https://www.ncbi.nlm.nih.gov/pubmed/21865176
http://dx.doi.org/10.1093/jxb/err235
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author Fojtová, Miloslava
Peška, Vratislav
Dobšáková, Zuzana
Mozgová, Iva
Fajkus, Jiří
Sýkorová, Eva
author_facet Fojtová, Miloslava
Peška, Vratislav
Dobšáková, Zuzana
Mozgová, Iva
Fajkus, Jiří
Sýkorová, Eva
author_sort Fojtová, Miloslava
collection PubMed
description Analysis of plants bearing a T-DNA insertion is a potent tool of modern molecular biology, providing valuable information about the function and involvement of genes in metabolic pathways. A collection of 12 Arabidopsis thaliana lines with T-DNA insertions in the gene coding for the catalytic subunit of telomerase (AtTERT) and in adjacent regions was screened for telomerase activity [telomere repeat amplification protocol (TRAP) assay], telomere length (terminal restriction fragments), and AtTERT transcription (quantitative reverse transcription-PCR). Lines with the insertion located upstream of the start codon displayed unchanged telomere stability and telomerase activity, defining a putative minimal AtTERT promoter and the presence of a regulatory element linked to increased transcription in the line SALK_048471. Lines bearing a T-DNA insertion inside the protein-coding region showed telomere shortening and lack of telomerase activity. Transcription in most of these lines was unchanged upstream of the T-DNA insertion, while it was notably decreased downstream. The expression profile varied markedly in mutant lines harbouring insertions at the 5' end of AtTERT which showed increased transcription and abolished tissue specificity. Moreover, the line FLAG_385G01 (T-DNA insertion inside intron 1) revealed the presence of a highly abundant downstream transcript with normal splicing but without active telomerase. The role of regulatory elements found along the AtTERT gene is discussed in respect to natural telomerase expression and putative intron-mediated enhancement.
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spelling pubmed-32230502011-11-23 Molecular analysis of T-DNA insertion mutants identified putative regulatory elements in the AtTERT gene Fojtová, Miloslava Peška, Vratislav Dobšáková, Zuzana Mozgová, Iva Fajkus, Jiří Sýkorová, Eva J Exp Bot Research Papers Analysis of plants bearing a T-DNA insertion is a potent tool of modern molecular biology, providing valuable information about the function and involvement of genes in metabolic pathways. A collection of 12 Arabidopsis thaliana lines with T-DNA insertions in the gene coding for the catalytic subunit of telomerase (AtTERT) and in adjacent regions was screened for telomerase activity [telomere repeat amplification protocol (TRAP) assay], telomere length (terminal restriction fragments), and AtTERT transcription (quantitative reverse transcription-PCR). Lines with the insertion located upstream of the start codon displayed unchanged telomere stability and telomerase activity, defining a putative minimal AtTERT promoter and the presence of a regulatory element linked to increased transcription in the line SALK_048471. Lines bearing a T-DNA insertion inside the protein-coding region showed telomere shortening and lack of telomerase activity. Transcription in most of these lines was unchanged upstream of the T-DNA insertion, while it was notably decreased downstream. The expression profile varied markedly in mutant lines harbouring insertions at the 5' end of AtTERT which showed increased transcription and abolished tissue specificity. Moreover, the line FLAG_385G01 (T-DNA insertion inside intron 1) revealed the presence of a highly abundant downstream transcript with normal splicing but without active telomerase. The role of regulatory elements found along the AtTERT gene is discussed in respect to natural telomerase expression and putative intron-mediated enhancement. Oxford University Press 2011-11 2011-08-23 /pmc/articles/PMC3223050/ /pubmed/21865176 http://dx.doi.org/10.1093/jxb/err235 Text en © 2011 The Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)
spellingShingle Research Papers
Fojtová, Miloslava
Peška, Vratislav
Dobšáková, Zuzana
Mozgová, Iva
Fajkus, Jiří
Sýkorová, Eva
Molecular analysis of T-DNA insertion mutants identified putative regulatory elements in the AtTERT gene
title Molecular analysis of T-DNA insertion mutants identified putative regulatory elements in the AtTERT gene
title_full Molecular analysis of T-DNA insertion mutants identified putative regulatory elements in the AtTERT gene
title_fullStr Molecular analysis of T-DNA insertion mutants identified putative regulatory elements in the AtTERT gene
title_full_unstemmed Molecular analysis of T-DNA insertion mutants identified putative regulatory elements in the AtTERT gene
title_short Molecular analysis of T-DNA insertion mutants identified putative regulatory elements in the AtTERT gene
title_sort molecular analysis of t-dna insertion mutants identified putative regulatory elements in the attert gene
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3223050/
https://www.ncbi.nlm.nih.gov/pubmed/21865176
http://dx.doi.org/10.1093/jxb/err235
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