Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia

Detalhes bibliográficos
Autor(a) principal: Santos, Eduardo José dos
Data de Publicação: 2019
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFG
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/10010
Resumo: The Trypanosoma cruzi, etiologic agent of Chagas disease is a flagellate protozoan transmitted by insect bugs (Reduvidae:Triatomine) to humans and wild mammals. The T. cruzi contains DNA in the nucleus (nDNA) and in the mitochondrion (kDNA). The kinetoplastid protozoan comprises maxicircles and and minicircles kDNA network sequences that integrate in the host’s cell genome. Insertional integrations of kDNA sequences bear high affinity to retrotransposable LINE-1 randomly distributed in the genome of somatic and germ line cells, and, thus generate an increasing diversity due to recombination and reshuffling. However, the profiles of kDNA integrations into the human genome in different Brazilian Ecosystems are unknown. In this regard, the ratio of transfer of T. cruzi minicircles sequences into the human genome was south. The study consisted of analyses of 36 members of two families from São Felipe-Bahia where Chagas disease is endemic. The study showed that 22% (8/36) families’ members had the diagnoses of the disease by immunofluorescence (IF) and enzyme-linked immunosorbant assay (ELISA). Yet, the PCR assays showed 61% (22/36) of families’ members had kDNA and nDNA assays positive for the T. cruzi infection. In order to detect the kDNA integration sites in the human genome, a tpTAIL-PCR (Targeted Primer Thermal Assymetric Interlaced-PCR), which consisted in combination of primers derived from T. cruzi minicircle conserved sequence with human LINE-1 sequence derived primers, was employed. The amplicon sequences obtained were subjected to homology analyses in Blast software. The hybrid kDNA-human DNA sequences revealed lateral and vertical transfer of T. cruzi minicircles into the genome of Chagas parents and progeny. Unravel of dynamics of mutations contributes to define variable profiles of kDNA minicircle sequence insertions into the human genome at different Ecosystems.
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spelling Lacerda, Elisângela de Paula Silveirahttp://lattes.cnpq.br/9390789693192751Cardoso, Clever Gomeshttp://lattes.cnpq.br/9545455455623006Teixeira, Antônio Raimundo Lima CruzZapata, Marco Túlio Antônio Garciahttp://lattes.cnpq.br/2863457276816692Santos, Eduardo José dos2019-09-11T15:36:04Z2019-02-26SANTOS, E. J. Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia. 2019. 106 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2019.http://repositorio.bc.ufg.br/tede/handle/tede/10010The Trypanosoma cruzi, etiologic agent of Chagas disease is a flagellate protozoan transmitted by insect bugs (Reduvidae:Triatomine) to humans and wild mammals. The T. cruzi contains DNA in the nucleus (nDNA) and in the mitochondrion (kDNA). The kinetoplastid protozoan comprises maxicircles and and minicircles kDNA network sequences that integrate in the host’s cell genome. Insertional integrations of kDNA sequences bear high affinity to retrotransposable LINE-1 randomly distributed in the genome of somatic and germ line cells, and, thus generate an increasing diversity due to recombination and reshuffling. However, the profiles of kDNA integrations into the human genome in different Brazilian Ecosystems are unknown. In this regard, the ratio of transfer of T. cruzi minicircles sequences into the human genome was south. The study consisted of analyses of 36 members of two families from São Felipe-Bahia where Chagas disease is endemic. The study showed that 22% (8/36) families’ members had the diagnoses of the disease by immunofluorescence (IF) and enzyme-linked immunosorbant assay (ELISA). Yet, the PCR assays showed 61% (22/36) of families’ members had kDNA and nDNA assays positive for the T. cruzi infection. In order to detect the kDNA integration sites in the human genome, a tpTAIL-PCR (Targeted Primer Thermal Assymetric Interlaced-PCR), which consisted in combination of primers derived from T. cruzi minicircle conserved sequence with human LINE-1 sequence derived primers, was employed. The amplicon sequences obtained were subjected to homology analyses in Blast software. The hybrid kDNA-human DNA sequences revealed lateral and vertical transfer of T. cruzi minicircles into the genome of Chagas parents and progeny. Unravel of dynamics of mutations contributes to define variable profiles of kDNA minicircle sequence insertions into the human genome at different Ecosystems.O Trypanosoma cruzi, agente etiológico da doença de Chagas, é um protozoário flagelado que pode ser transmitido por insetos hematófagos da família Reduviidae. O T. cruzi possui DNA no núcleo (nDNA) e na mitocôndria (kDNA). O kDNA é composto de maxicírculos e minicírculos concatenados em uma extensa rede involucrada pelo cinetoplasto. Sequências de minicírculos de kDNA podem se inserir no genoma de célula hospedeira. A inserção de sequências de minicírculos ocorre com maior frequência em retrotransposons LINE-1, tem distribuição randômica em células somáticas e germinativas de parentais e progênies, e aumenta a diversidade mediante rearranjos e recombinação nos sítios de integração. Contudo, o perfil de integração de minicírculos de kDNA em chagásicos de diferentes regiões do Brasil ainda é pouco conhecido. A análise da transferência lateral e vertical de minicírculos de kDNA em 36 indivíduos de duas famílias de São Felipe, Estado da Bahia revelou que 22% (8/36) dos investigados tiveram diagnóstico de doença de Chagas pelos testes imunofluorescência indireta (IF) e imunoensaio enzymático (ELISA). Todavia, os testes de ácidos nucleicos (PCR) mostraram que 61% (22/36) dos investigados foram positivos para kDNA e nDNA, e, portanto, eram portadores da doença de Chagas. Em seguida, foi utilizada uma metodologia com base na combinação de iniciadores de região conservada de minicírculos de kDNA de T. cruzi e de sequência de DNA humano (tpTAIL-PCR: Targeted Primer Thermal Assymetric Interlaced-PCR), para identificar os sítios de integração do kDNA. Os amplicons foram sequenciados e submetidos à análise de homologia pelo software Blast. Os resultados comprovam transferência horizontal e vertical de sequências de minicírculos de kDNA no genoma do chagásico. O entendimento da dinâmica dessas mutações contribui para a identificação de padrões de inserções de sequências de minicírculos de kDNA no genoma de indivíduos de diferentes EcossistemasSubmitted by Franciele Moreira (francielemoreyra@gmail.com) on 2019-09-11T15:27:02Z No. of bitstreams: 2 Dissertação - Eduardo José dos Santos - 2019.pdf: 4346886 bytes, checksum: d8a881b9ddc5408ae1dd36ccc508a9a4 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2019-09-11T15:36:03Z (GMT) No. of bitstreams: 2 Dissertação - Eduardo José dos Santos - 2019.pdf: 4346886 bytes, checksum: d8a881b9ddc5408ae1dd36ccc508a9a4 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2019-09-11T15:36:04Z (GMT). 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dc.title.eng.fl_str_mv Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia
title Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia
spellingShingle Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia
Santos, Eduardo José dos
Trypanosoma cruzi
kDNA
Doença de Chagas
Retrotransposons
Trypanosoma cruzi
Chagas disease
BIOQUIMICA::BIOLOGIA MOLECULAR
title_short Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia
title_full Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia
title_fullStr Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia
title_full_unstemmed Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia
title_sort Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia
author Santos, Eduardo José dos
author_facet Santos, Eduardo José dos
author_role author
dc.contributor.advisor1.fl_str_mv Lacerda, Elisângela de Paula Silveira
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/9390789693192751
dc.contributor.advisor-co1.fl_str_mv Cardoso, Clever Gomes
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/9545455455623006
dc.contributor.referee1.fl_str_mv Teixeira, Antônio Raimundo Lima Cruz
dc.contributor.referee2.fl_str_mv Zapata, Marco Túlio Antônio Garcia
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/2863457276816692
dc.contributor.author.fl_str_mv Santos, Eduardo José dos
contributor_str_mv Lacerda, Elisângela de Paula Silveira
Cardoso, Clever Gomes
Teixeira, Antônio Raimundo Lima Cruz
Zapata, Marco Túlio Antônio Garcia
dc.subject.por.fl_str_mv Trypanosoma cruzi
kDNA
Doença de Chagas
Retrotransposons
topic Trypanosoma cruzi
kDNA
Doença de Chagas
Retrotransposons
Trypanosoma cruzi
Chagas disease
BIOQUIMICA::BIOLOGIA MOLECULAR
dc.subject.eng.fl_str_mv Trypanosoma cruzi
Chagas disease
dc.subject.cnpq.fl_str_mv BIOQUIMICA::BIOLOGIA MOLECULAR
description The Trypanosoma cruzi, etiologic agent of Chagas disease is a flagellate protozoan transmitted by insect bugs (Reduvidae:Triatomine) to humans and wild mammals. The T. cruzi contains DNA in the nucleus (nDNA) and in the mitochondrion (kDNA). The kinetoplastid protozoan comprises maxicircles and and minicircles kDNA network sequences that integrate in the host’s cell genome. Insertional integrations of kDNA sequences bear high affinity to retrotransposable LINE-1 randomly distributed in the genome of somatic and germ line cells, and, thus generate an increasing diversity due to recombination and reshuffling. However, the profiles of kDNA integrations into the human genome in different Brazilian Ecosystems are unknown. In this regard, the ratio of transfer of T. cruzi minicircles sequences into the human genome was south. The study consisted of analyses of 36 members of two families from São Felipe-Bahia where Chagas disease is endemic. The study showed that 22% (8/36) families’ members had the diagnoses of the disease by immunofluorescence (IF) and enzyme-linked immunosorbant assay (ELISA). Yet, the PCR assays showed 61% (22/36) of families’ members had kDNA and nDNA assays positive for the T. cruzi infection. In order to detect the kDNA integration sites in the human genome, a tpTAIL-PCR (Targeted Primer Thermal Assymetric Interlaced-PCR), which consisted in combination of primers derived from T. cruzi minicircle conserved sequence with human LINE-1 sequence derived primers, was employed. The amplicon sequences obtained were subjected to homology analyses in Blast software. The hybrid kDNA-human DNA sequences revealed lateral and vertical transfer of T. cruzi minicircles into the genome of Chagas parents and progeny. Unravel of dynamics of mutations contributes to define variable profiles of kDNA minicircle sequence insertions into the human genome at different Ecosystems.
publishDate 2019
dc.date.accessioned.fl_str_mv 2019-09-11T15:36:04Z
dc.date.issued.fl_str_mv 2019-02-26
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dc.identifier.citation.fl_str_mv SANTOS, E. J. Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia. 2019. 106 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2019.
dc.identifier.uri.fl_str_mv http://repositorio.bc.ufg.br/tede/handle/tede/10010
identifier_str_mv SANTOS, E. J. Análise da herança do DNA do Trypanosoma cruzi em parentais e progênies de chagásicos de famílias de São Felipe – Bahia. 2019. 106 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2019.
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