Análises moleculares na subfamília hypoptopomatinae (Siluriformes, Loricariidae): isolamento, caracterização e mapeamento cromossômico de sequências repetitivas
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2009-06-05
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Ferreira, Daniela Cristina [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Uma grande fração do genoma dos eucariotos é constituída de sequências repetitivas, que estão divididas em duas classes. Na primeira classe estão as sequências repetidas em cadeia, (satélites, minissatélites e microssatélites) e na segunda estão as sequências repetidas dispersas, que podem ser divididas em transposons e retrotransposons. Poucos estudos têm sido realizados acerca dessas sequências para elucidar sua função no genoma, que até pouco tempo eram conhecidas como DNA egoísta ou lixo. Porém, estudos recentes tem revelado que essas sequências estão envolvidas na diferenciação cromossômicas sexuais e na organização funcional e estrutural do genoma, além dessas sequências repetitivas constituírem bons marcadores cromossômicos. A subfamília Hypoptopomatina é caracterizada citogeneticamente por apresentar-se conservada quando ao número diplóide, que é de 2n=54 cromossomos para a maioria das espécies analisadas e uma distribuição da hetecocromatina bastante heterogênea, o que torna as espécies dessa subfamília bastante interessantes para os estudos das sequências repetidas, uma vez que essas sequências se localizam preferencialmente em regiões heterocromáticas. No presente trabalho foi estudada a localização de sequências repetidas dispersas Rex1, Rex3 via PCR em quatorze espécies da subfamília Hypoptopomatinae e do elemento disperso HLBa isolado do genoma de Hisonotus leucofrenatus por digestão enzimática. Os elementos Rex1 e Rex3 apresentaram-se conservados no genoma da subfamília Hypoptopomatinae, bem como em espécies distantes filogeneticamente. Tanto Rex1 e Rex3 quanto HLBam se encontram dispersos no genoma, porém este forma blocos em regiões heterocromáticas. Além disso, foi isolado o satélite PTHind do genoma de Pseudotocinclus tientensis, que encontra-se exclusivamente no genoma da espécie isolada, constituindo...
A great fraction of eukaryotic genomic is based on repeated sequences, which are divided in two classes. The first are in tandem (satellites , minisatellites and micro satellites and in the second class are diverse repeated sequences, that can be divided in transposons and retrotransposons. Only few studies had been conducted with these sequences to bring into the light its genomic function, known by the moment as selfish DNA or junk DNA. However, the few researches conducted may indicate that these sequences are related to sexual chromosome deferential and functional and structural genomic organization. In addition, repeated sequences can be used as effective chromosome markers. The subfamily Hypoptopomatinae's cytogenetic characteristics are the conservation of diploid number, which are 2n=54 chromosomes and heterogenic heterochromatic distribution. Based on that, the species of this subfamily became very interesting for analyze and study repeated sequences and also by the fact of these sequences are mostly located in heterochromatic regions. In the present study, Rex1 and Rex3 dispersed repeated sequences were isolated by PCR in fourteen species of Hypoptomatinae subfamily and the dispersed HLBam element was isolated in Hisonotus leucofrenatus genomic by enzymatic digestion. The Rex1 and Rex3 elements are conserved in Hypoptopomatinae subfamily genomic, likewise in phylogenetic distant species. Either Rex1, Rex3 or HLBam are dispersed in genomic, however, in block form inside heterochromatic regions. Moreover, the PTHind satellite was isolated from Pseudotocinclus tientensis genomic, which is exclusively encountered in the genomic of this isolate specie and may be a good marker for the specie. Additionally, sex-specific sequences were isolated by the AFLP technique from the genomic of Hisonotus leucofrenatus females. In Hypoptopomatinae subfamily the major part of current... (Complete abstract click electronic access below)
A great fraction of eukaryotic genomic is based on repeated sequences, which are divided in two classes. The first are in tandem (satellites , minisatellites and micro satellites and in the second class are diverse repeated sequences, that can be divided in transposons and retrotransposons. Only few studies had been conducted with these sequences to bring into the light its genomic function, known by the moment as selfish DNA or junk DNA. However, the few researches conducted may indicate that these sequences are related to sexual chromosome deferential and functional and structural genomic organization. In addition, repeated sequences can be used as effective chromosome markers. The subfamily Hypoptopomatinae's cytogenetic characteristics are the conservation of diploid number, which are 2n=54 chromosomes and heterogenic heterochromatic distribution. Based on that, the species of this subfamily became very interesting for analyze and study repeated sequences and also by the fact of these sequences are mostly located in heterochromatic regions. In the present study, Rex1 and Rex3 dispersed repeated sequences were isolated by PCR in fourteen species of Hypoptomatinae subfamily and the dispersed HLBam element was isolated in Hisonotus leucofrenatus genomic by enzymatic digestion. The Rex1 and Rex3 elements are conserved in Hypoptopomatinae subfamily genomic, likewise in phylogenetic distant species. Either Rex1, Rex3 or HLBam are dispersed in genomic, however, in block form inside heterochromatic regions. Moreover, the PTHind satellite was isolated from Pseudotocinclus tientensis genomic, which is exclusively encountered in the genomic of this isolate specie and may be a good marker for the specie. Additionally, sex-specific sequences were isolated by the AFLP technique from the genomic of Hisonotus leucofrenatus females. In Hypoptopomatinae subfamily the major part of current... (Complete abstract click electronic access below)
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Peixe - Genética molecular, Hypoptopomatinae
Como citar
FERREIRA, Daniela Cristina. Análises moleculares na subfamília hypoptopomatinae (Siluriformes, Loricariidae): isolamento, caracterização e mapeamento cromossômico de sequências repetitivas. 2009. 146 f. Tese (doutorado) - Universidade Estadual Paulista, Instituto de Biociências de Botucatu, 2009.