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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2007.tde-28012008-120815
Document
Author
Full name
Marinalva Martins Pinheiro
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2007
Supervisor
Committee
Menck, Carlos Frederico Martins (President)
Barros, Mário Henrique de
Oliveira, Carla Columbano de
Oliveira, Mariana Cabral de
Schenberg, Ana Clara Guerrini
Title in Portuguese
Busca de genes relacionados a fenótipos mutadores em Caulobacter crescentus.
Keywords in Portuguese
Caulobacter crescentus
Genômica.
Lesão do DNA
Mutadores
Mutagênese
Reparo de DNA
Abstract in Portuguese
A comparação "in silico" das vias de reparo de DNA nos genomas de Caulobacter crescentus e E. coli mostra diferenças significativas entre estas duas bactérias, sugerindo diversidade biológica das respostas a danos no DNA entre as bactérias. Em busca de genes que possam proteger o genoma de C. crescentus contra mutações, foi feita uma varredura em uma biblioteca de cerca de 5.000 clones construída a partir de inserção aleatória do transposon TN5 no genoma dessa bactéria. A maioria destes genes não tinha sido, ainda, relacionada a fenótipo mutador, mas, como esperado, também identificamos genes já conhecidos como relacionados a fenótipos mutadores. Alguns clones candidatos foram investigados quanto ao tipo de mutação induzida, baseando-se na sequência do gene rpoB, com o objetivo de indicar o processo mutacional sob condições genéticas específicas. Análises baseadas na medida da atividade promotora em fusão de transcrição com lacZ, mostra que o sistema SOS está alterado em alguns clones, e pode justificar parte do fenótipo mutador em alguns deles.
Title in English
Search for genes related to the mutator phenotypes in Caulobacter crescentus.
Keywords in English
Caulobacter crescentus
DNA damage
DNA repair
Genomic.
Mutagênesis
Mutators
Abstract in English
The "in silico" comparison of the main DNA repair genes between C. crescentus and E. coli shows significant differences between these bacteria, suggesting biological diversity in bacterial responses to DNA damage. We further screened a C. crescentus library of 5,000 clones mutated by random insertion of the TN5 transposon in the genome, searching for clones with high levels of spontaneous rifampicin resistance mutations. Most of the genes identified have not been previously reported as related to mutator phenotype, but, as expected, we have also identified proteins already known as causing mutator phenotypes when mutated. As part of the functional characterization, some candidate clones were characterized based on rpoB gene sequences aiming at indicating the mutational process under specific genetic background conditions. Analysis of some candidate clones based on measurements of promoter activity with lacZ transcriptional fusions show that the system SOS is modified in some clones and this justify part of their mutator phenotype.
 
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Publishing Date
2008-02-13
 
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