Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua)

Detalhes bibliográficos
Autor(a) principal: Szrajer, Marcos Rosolino
Data de Publicação: 2011
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFSCAR
Texto Completo: https://repositorio.ufscar.br/handle/ufscar/8796
Resumo: The fruit flies (Anastrepha sp) are organisms endemic to the Neotropics that are similar morphologically but present some behavioral, reproductive and molecular variations. Among these insects we can highlight the fraterculus group, which consists of a set of cryptic species, between them Anastrepha oblique, that represents threats to national fruit culture. The effective control of this grouping requires the expansion of the few studies concerning to the biology and history of populations which has a very complex systematic. Previous works suggest a high rate of evolution of genes related to reproductive issues and recognition of partners due to the high selective pressures involved in that relationship, hence, we chose to work with the fruitless gene which is recognized for its role in sexual differentiation and male courtship behavior in dipteran insects. Since a small region of this gene was already been amplified in our laboratory, we amplified and described the flanking sequences by RACE-PCR technique which used primers constructed targeting the 5 'and 3' ends. We used cDNA libraries produced from the extraction of mRNAs from different stages of development of these organisms enabling the identification of different isoforms arising from alternative splicing and structural characterization of the coding region (CDS) common to both sexes. We compared the data generated with the corresponding sequences of the fruitless gene in other insect species identifying the rate of similarity for nucleotides and amino acids (identity ratio) which revealed that the éxons coding for specific domains such as BTB and Zinc Fingers tend to remain more conserved in relation to other regions and éxons from Connector Region. Subsequently, we performed positive selection tests through the Relaxed Branch-site test and Strict Branch-site test, which were contrasted by Likelihood Ratio Tests (LRT) which identified 25 new sites evolving under positive selection, corroborating the findings of Sobrinho e de Brito (2010) and suggesting the presence of these forces on specific regions of this important gene linked to reproductive aspects of several major groups of insects.
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spelling Szrajer, Marcos RosolinoBrito, Reinaldo Alves dehttp://lattes.cnpq.br/8253066295947754http://lattes.cnpq.br/990008523357049487b7ab20-b0bc-4dec-a4bd-b5a58937f6bf2017-06-01T13:02:45Z2017-06-01T13:02:45Z2011-10-28SZRAJER, Marcos Rosolino. Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua). 2011. Dissertação (Mestrado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2011. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8796.https://repositorio.ufscar.br/handle/ufscar/8796The fruit flies (Anastrepha sp) are organisms endemic to the Neotropics that are similar morphologically but present some behavioral, reproductive and molecular variations. Among these insects we can highlight the fraterculus group, which consists of a set of cryptic species, between them Anastrepha oblique, that represents threats to national fruit culture. The effective control of this grouping requires the expansion of the few studies concerning to the biology and history of populations which has a very complex systematic. Previous works suggest a high rate of evolution of genes related to reproductive issues and recognition of partners due to the high selective pressures involved in that relationship, hence, we chose to work with the fruitless gene which is recognized for its role in sexual differentiation and male courtship behavior in dipteran insects. Since a small region of this gene was already been amplified in our laboratory, we amplified and described the flanking sequences by RACE-PCR technique which used primers constructed targeting the 5 'and 3' ends. We used cDNA libraries produced from the extraction of mRNAs from different stages of development of these organisms enabling the identification of different isoforms arising from alternative splicing and structural characterization of the coding region (CDS) common to both sexes. We compared the data generated with the corresponding sequences of the fruitless gene in other insect species identifying the rate of similarity for nucleotides and amino acids (identity ratio) which revealed that the éxons coding for specific domains such as BTB and Zinc Fingers tend to remain more conserved in relation to other regions and éxons from Connector Region. Subsequently, we performed positive selection tests through the Relaxed Branch-site test and Strict Branch-site test, which were contrasted by Likelihood Ratio Tests (LRT) which identified 25 new sites evolving under positive selection, corroborating the findings of Sobrinho e de Brito (2010) and suggesting the presence of these forces on specific regions of this important gene linked to reproductive aspects of several major groups of insects.As moscas-das-frutas (Anastrepha sp) são organismos endêmicos da região neotropical que apesar de serem semelhantes morfologicamente distinguem-se por variações comportamentais, reprodutivas e moleculares. Entre esses insetos podemos destacar o grupo fraterculus o qual é composto por um conjunto de espécies crípticas, entre elas Anastrepha obliqua, que representam ameaças à fruticultura nacional. O controle eficiente deste agrupamento exige a ampliação dos ainda escassos estudos referentes à biologia e história das populações cuja sistemática tem se mostrado bastante complexa. Trabalhos anteriores sugerem uma alta taxa evolutiva agindo sobre genes relacionados a aspectos reprodutivos e reconhecimento de parceiros devido as elevadas pressões seletivas envolvidas nessa relação, para tanto, escolhemos trabalhar com o gene fruitless por sua reconhecida importância na diferenciação sexual e comportamento de corte nos machos nos insetos dípteros. Uma vez que dispúnhamos de uma pequena região já amplificada deste gene em nosso laboratório, amplificamos as sequências flanqueadoras através da técnica de RACE-PCR que utilizou primers construídos e orientados para as extremidades 5´ e 3´. Foram utilizadas bibliotecas de cDNAs produzidas a partir da extração de mRNAs de diferentes estágios de desenvolvimento desses organismos possibilitando a identificação de diferentes isoformas provenientes de splincing alternativo e caracterização estrutural da região codificadora (CDS) comum a ambos os sexos. Comparamos os dados gerados com as sequencias correspondentes do gene fruitless em outras espécies de insetos identificando sua taxa de identidade nucleotídica e de aminoácidos as quais revelaram que os éxons que codificam domínios específicos como BTB e Zinc Fingers tendem a se manter mais conservados em relação às demais regiões e aos éxons da Região Conectora. Posteriormente, realizamos testes de seleção positiva conhecidos como Relaxed Branch-site test e Strict Branch-site test, os quais foram contrastados pelo Likelihood Ratio Test e possibilitaram a identificação de 25 novos sítios evoluindo sob seleção positiva, corroborando as descobertas de Sobrinho e de Brito (2010) e sugerindo a atuação dessas forças sobre regiões específicas deste importante gene ligado aos aspectos reprodutivos de diversos grupos principais de insetos.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEvUFSCarGenéticaMoscas-das-frutasAnastrepha spFraterculus groupFruitlessCIENCIAS BIOLOGICAS::GENETICACaracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisOnline600600e4ebf16c-a933-4cae-b428-d05e5cc0dc96info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARLICENSElicense.txtlicense.txttext/plain; charset=utf-81957https://repositorio.ufscar.br/bitstream/ufscar/8796/2/license.txtae0398b6f8b235e40ad82cba6c50031dMD52ORIGINALDissMRS.pdfDissMRS.pdfapplication/pdf3014427https://repositorio.ufscar.br/bitstream/ufscar/8796/3/DissMRS.pdf34444caee9bbab6499092a2cd3942b2eMD53TEXTDissMRS.pdf.txtDissMRS.pdf.txtExtracted texttext/plain164520https://repositorio.ufscar.br/bitstream/ufscar/8796/4/DissMRS.pdf.txt4cb7da2a5ac6e1b793de6a6bd0b96631MD54THUMBNAILDissMRS.pdf.jpgDissMRS.pdf.jpgIM Thumbnailimage/jpeg5275https://repositorio.ufscar.br/bitstream/ufscar/8796/5/DissMRS.pdf.jpg88f33dcad6f4f6647d1cb7b3ea49c554MD55ufscar/87962023-09-18 18:31:24.359oai:repositorio.ufscar.br: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Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:24Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false
dc.title.por.fl_str_mv Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua)
title Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua)
spellingShingle Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua)
Szrajer, Marcos Rosolino
Genética
Moscas-das-frutas
Anastrepha sp
Fraterculus group
Fruitless
CIENCIAS BIOLOGICAS::GENETICA
title_short Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua)
title_full Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua)
title_fullStr Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua)
title_full_unstemmed Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua)
title_sort Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua)
author Szrajer, Marcos Rosolino
author_facet Szrajer, Marcos Rosolino
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/9900085233570494
dc.contributor.author.fl_str_mv Szrajer, Marcos Rosolino
dc.contributor.advisor1.fl_str_mv Brito, Reinaldo Alves de
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/8253066295947754
dc.contributor.authorID.fl_str_mv 87b7ab20-b0bc-4dec-a4bd-b5a58937f6bf
contributor_str_mv Brito, Reinaldo Alves de
dc.subject.por.fl_str_mv Genética
Moscas-das-frutas
topic Genética
Moscas-das-frutas
Anastrepha sp
Fraterculus group
Fruitless
CIENCIAS BIOLOGICAS::GENETICA
dc.subject.lat.fl_str_mv Anastrepha sp
dc.subject.eng.fl_str_mv Fraterculus group
Fruitless
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS::GENETICA
description The fruit flies (Anastrepha sp) are organisms endemic to the Neotropics that are similar morphologically but present some behavioral, reproductive and molecular variations. Among these insects we can highlight the fraterculus group, which consists of a set of cryptic species, between them Anastrepha oblique, that represents threats to national fruit culture. The effective control of this grouping requires the expansion of the few studies concerning to the biology and history of populations which has a very complex systematic. Previous works suggest a high rate of evolution of genes related to reproductive issues and recognition of partners due to the high selective pressures involved in that relationship, hence, we chose to work with the fruitless gene which is recognized for its role in sexual differentiation and male courtship behavior in dipteran insects. Since a small region of this gene was already been amplified in our laboratory, we amplified and described the flanking sequences by RACE-PCR technique which used primers constructed targeting the 5 'and 3' ends. We used cDNA libraries produced from the extraction of mRNAs from different stages of development of these organisms enabling the identification of different isoforms arising from alternative splicing and structural characterization of the coding region (CDS) common to both sexes. We compared the data generated with the corresponding sequences of the fruitless gene in other insect species identifying the rate of similarity for nucleotides and amino acids (identity ratio) which revealed that the éxons coding for specific domains such as BTB and Zinc Fingers tend to remain more conserved in relation to other regions and éxons from Connector Region. Subsequently, we performed positive selection tests through the Relaxed Branch-site test and Strict Branch-site test, which were contrasted by Likelihood Ratio Tests (LRT) which identified 25 new sites evolving under positive selection, corroborating the findings of Sobrinho e de Brito (2010) and suggesting the presence of these forces on specific regions of this important gene linked to reproductive aspects of several major groups of insects.
publishDate 2011
dc.date.issued.fl_str_mv 2011-10-28
dc.date.accessioned.fl_str_mv 2017-06-01T13:02:45Z
dc.date.available.fl_str_mv 2017-06-01T13:02:45Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv SZRAJER, Marcos Rosolino. Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua). 2011. Dissertação (Mestrado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2011. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8796.
dc.identifier.uri.fl_str_mv https://repositorio.ufscar.br/handle/ufscar/8796
identifier_str_mv SZRAJER, Marcos Rosolino. Caracterização e análise da região codificadora do gene fruitless em moscas-das-frutas (Anastrepha obliqua). 2011. Dissertação (Mestrado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2011. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8796.
url https://repositorio.ufscar.br/handle/ufscar/8796
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language por
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dc.publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEv
dc.publisher.initials.fl_str_mv UFSCar
publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
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