Desenvolvimento de marcadores SNPs (Single Nucleotide Polymorphisms) para estudos populacionais do mosquito Aedes aegypti no Brasil
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Data
2008-08-21
Autores
Paduan, Karina dos Santos [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Aedes aegypti é o principal vetor dos vírus dengue e representa cada vez mais um problema de saúde pública. O uso de inseticidas de origem orgânica ou inorgânica é uma das metodologias mais adotadas como parte do controle de vetores, entretanto, processos de resistência têm sido detectados freqüentemente. O desenvolvimento de ferramentas moleculares tem facilitado o entendimento das relações entre o vetor, patógeno e o homem. Os marcadores genéticos mais utilizados em estudos de genética de populações geralmente representam um trabalho laboratorial intenso e, em alguns casos, são limitados pela quantidade de DNA genômico extraído por indivíduo. Recentemente, a tecnologia disponível, permitiu o desenvolvimento dos marcadores SNPs (Single Nucleotide Polymorphisms) que correspondem a diferenças de um único nucleotídeo na seqüência de DNA. Assim, o presente trabalho objetivou desenvolver marcadores SNPs localizados em genes nucleares, do mosquito Ae. aegypti, para aplicação na genotipagem de populações deste vetor no Brasil. Para tanto, foram investigada seqüencias em dezoito genes nucleares de dezesseis populações do Ae. aegypti, e nove destes genes foram selecionados para a presença de SNPs. Os oito marcadores mais polimórficos foram utilizados na caracterização de três populações do mosquito anteriormente analisadas utilizando-se as técnicas de RAPD (Random Amplified Polymorphic DNA) e seqüenciamento de genes mitocondriais. Os resultados obtidos com os SNPs mostraram um alto grau de polimorfismo intra e inter populacional, mais significativo quando comparado com os estudos realizados anteriormente, além da presença de duas diferentes linhagens genéticas entre as populações geográficas estudadas.
Aedes aegypti is the primary vector of dengue fever virus and it represents a major problem of Public Health. The use of insecticides of organic or inorganic origin is one of the more accepted methodologies as part of this control, however, resistance processes have been detected constantly. The development of molecular tools has made possible the understanding of the relationships among vector, pathogen and man. The genetic markers more used in studies of genetics populations usually represent an intense laboratorial work and in some cases they are limited for the amount of genomic DNA extracted by individual. Recently, the available technology, allowed the development of the markers SNPs (Single Nucleotide Polymorphism) that correspond to differences of a single nucleotide in the sequence of DNA. Thus, this work aims to develop of SNPs markers in nuclear genes of the Ae. aegypti for application in genotyping of populations of this vector in Brazil. For so much, were investigated sequences in eighteen nuclear genes of sixteen populations of Ae. aegypti, and nine of this genes were selected to presence of the SNPs. Eight more polymorphic markers were used in the characterization of three populations of the mosquito previously analyzed using the techniques of RAPD (Random Amplified Polymorphic DNA) and sequencing of mitochondrial genes. Results obtained with the SNPs showed a high degree of polymorphic intra and interpopulations, more significantly when compared with the prior studies, besides the presence of two different genetic lineages distributed among the studied geographical populations.
Aedes aegypti is the primary vector of dengue fever virus and it represents a major problem of Public Health. The use of insecticides of organic or inorganic origin is one of the more accepted methodologies as part of this control, however, resistance processes have been detected constantly. The development of molecular tools has made possible the understanding of the relationships among vector, pathogen and man. The genetic markers more used in studies of genetics populations usually represent an intense laboratorial work and in some cases they are limited for the amount of genomic DNA extracted by individual. Recently, the available technology, allowed the development of the markers SNPs (Single Nucleotide Polymorphism) that correspond to differences of a single nucleotide in the sequence of DNA. Thus, this work aims to develop of SNPs markers in nuclear genes of the Ae. aegypti for application in genotyping of populations of this vector in Brazil. For so much, were investigated sequences in eighteen nuclear genes of sixteen populations of Ae. aegypti, and nine of this genes were selected to presence of the SNPs. Eight more polymorphic markers were used in the characterization of three populations of the mosquito previously analyzed using the techniques of RAPD (Random Amplified Polymorphic DNA) and sequencing of mitochondrial genes. Results obtained with the SNPs showed a high degree of polymorphic intra and interpopulations, more significantly when compared with the prior studies, besides the presence of two different genetic lineages distributed among the studied geographical populations.
Descrição
Palavras-chave
Aedes aegypti - Genética, Mosquitos, Genes nucleares, Aedes aegypti, Dengue, Polymorphism
Como citar
PADUAN, Karina dos Santos. Desenvolvimento de marcadores SNPs (Single Nucleotide Polymorphisms) para estudos populacionais do mosquito Aedes aegypti no Brasil. 2008. 105 f. Tese (doutorado) - Universidade Estadual Paulista, Instituto de Biociências de Botucatu, 2008.