Methylation-specific PCR analysis in Col8A1 promoter in Creole cattle carrier of rob(1;29)

Robertsonian translocation (rob(1;29)) is the most frequent structural chromosomal abnormality in cattle. Heterozygous carriers have a normal phenotype but show a 3-5% decrease in fertility. Chromatin decondensation was evaluated similar to the inactive X chromosome when submitted to demethylating a...

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Autores principales:	Postiglioni,Alicia, García,Cristina B, Rincón,Gonzalo, Arruga,M. Victoria
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2011
Materias:	bisulfite collagen Creole cattle cytosines methylation
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000300013
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spelling	oai:scielo:S0717-345820110003000132011-09-23Methylation-specific PCR analysis in Col8A1 promoter in Creole cattle carrier of rob(1;29)Postiglioni,AliciaGarcía,Cristina BRincón,GonzaloArruga,M. Victoria bisulfite collagen Creole cattle cytosines methylation Robertsonian translocation (rob(1;29)) is the most frequent structural chromosomal abnormality in cattle. Heterozygous carriers have a normal phenotype but show a 3-5% decrease in fertility. Chromatin decondensation was evaluated similar to the inactive X chromosome when submitted to demethylating agent. Based on this result, and the concept that imprinted genes are essential in embryonic development, we decided to query genes located on BTA1 and BTA29 that could undergo genome imprinting. The collagen typeVIII-Î±1 (Col8A1) acted on extracellular matrix structural proteins. DNA bisulfite conversion and sequentiation methods were used to compare its differential methylation patterns. It was performed on eight Creole cattle DNA blood samples from normal and rob(1;29) carriers. An in silico screening for CpG islands in its promoter uncovered a single region of 454 bp prone to methylation. BiQ-Analizer software was used to show the selective conversion of unmethylated cytosines to uracils obtaining the following results: unmethylated CpGs: 0.000 (0 cases), methylated CpGs: 0.802 (77 cases) and CpGs not present: 0.198 (19 cases). No differences between samples were observed in this highly methylated region. This technique was successfully applied so it is a straightforward methodology that can be utilized to evaluate different tissue associated to specific gene expression.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.14 n.3 20112011-05-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000300013en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	bisulfite collagen Creole cattle cytosines methylation
spellingShingle	bisulfite collagen Creole cattle cytosines methylation Postiglioni,Alicia García,Cristina B Rincón,Gonzalo Arruga,M. Victoria Methylation-specific PCR analysis in Col8A1 promoter in Creole cattle carrier of rob(1;29)
description	Robertsonian translocation (rob(1;29)) is the most frequent structural chromosomal abnormality in cattle. Heterozygous carriers have a normal phenotype but show a 3-5% decrease in fertility. Chromatin decondensation was evaluated similar to the inactive X chromosome when submitted to demethylating agent. Based on this result, and the concept that imprinted genes are essential in embryonic development, we decided to query genes located on BTA1 and BTA29 that could undergo genome imprinting. The collagen typeVIII-Î±1 (Col8A1) acted on extracellular matrix structural proteins. DNA bisulfite conversion and sequentiation methods were used to compare its differential methylation patterns. It was performed on eight Creole cattle DNA blood samples from normal and rob(1;29) carriers. An in silico screening for CpG islands in its promoter uncovered a single region of 454 bp prone to methylation. BiQ-Analizer software was used to show the selective conversion of unmethylated cytosines to uracils obtaining the following results: unmethylated CpGs: 0.000 (0 cases), methylated CpGs: 0.802 (77 cases) and CpGs not present: 0.198 (19 cases). No differences between samples were observed in this highly methylated region. This technique was successfully applied so it is a straightforward methodology that can be utilized to evaluate different tissue associated to specific gene expression.
author	Postiglioni,Alicia García,Cristina B Rincón,Gonzalo Arruga,M. Victoria
author_facet	Postiglioni,Alicia García,Cristina B Rincón,Gonzalo Arruga,M. Victoria
author_sort	Postiglioni,Alicia
title	Methylation-specific PCR analysis in Col8A1 promoter in Creole cattle carrier of rob(1;29)
title_short	Methylation-specific PCR analysis in Col8A1 promoter in Creole cattle carrier of rob(1;29)
title_full	Methylation-specific PCR analysis in Col8A1 promoter in Creole cattle carrier of rob(1;29)
title_fullStr	Methylation-specific PCR analysis in Col8A1 promoter in Creole cattle carrier of rob(1;29)
title_full_unstemmed	Methylation-specific PCR analysis in Col8A1 promoter in Creole cattle carrier of rob(1;29)
title_sort	methylation-specific pcr analysis in col8a1 promoter in creole cattle carrier of rob(1;29)
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2011
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000300013
work_keys_str_mv	AT postiglionialicia methylationspecificpcranalysisincol8a1promoterincreolecattlecarrierofrob129 AT garciacristinab methylationspecificpcranalysisincol8a1promoterincreolecattlecarrierofrob129 AT rincongonzalo methylationspecificpcranalysisincol8a1promoterincreolecattlecarrierofrob129 AT arrugamvictoria methylationspecificpcranalysisincol8a1promoterincreolecattlecarrierofrob129
_version_	1718441839013396480

Methylation-specific PCR analysis in Col8A1 promoter in Creole cattle carrier of rob(1;29)

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