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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2013.tde-26062014-125358
Document
Author
Full name
Maristela Previato
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Marques, Marilis do Valle (President)
Gueiros Filho, Frederico José
Soares Netto, Luis Eduardo
Title in Portuguese
 Estudo da regulação de genes envolvidos na resposta a estresse oxidativo em Caulobacter crescentus.
Keywords in Portuguese
Bactérias gram negativas
Estresse oxidativo
Peroxidase
Regulação gênica
Superóxido dismutase
Abstract in Portuguese
O estresse oxidativo, causado por níveis aumentados de espécies reativas de oxigênio (ROS), pode causar danos celulares. Várias enzimas, como as subunidades da alquil-hidroperóxido redutase (AhpC e AhpF) e as superóxido dismutases (SOD), são responsáveis por remover as ROS. Os mecanismos de regulação da expressão gênica de C. crescentus para os genes ahpC, sodA, sodB e sodC, foram analisados com fusões de transcrição ao gene repórter lacZ, permitindo a quantificação da expressão por ensaios da atividade de b-galactosidase, e RT-PCR quantitativo. As culturas foram cultivadas em meio PYE ou M2 e a expressão de cada gene foi avaliada na presença de peróxido de hidrogênio (H2O2), tert-butil hidroperóxido (tBOOH), paraquat, menadiona, pirogalol, FeSO4 ou DDPi. Em C. crescentus ahpC é induzido por peróxidos e regulado por OxyR. As SODs são induzidas principalmente por superóxidos, sodB e sodC possuem indução de fase na fase estacionária e possivelmente estão sob o controle do sigma J, enquanto o gene sodA é regulado por sigma F e sigma J.
Title in English
Regulation of genes involved in oxidative stress response in Caulobacter crescentus.
Keywords in English
Gene regulation
Gram-negative bacteria
Oxidative stress
Peroxidase
Superoxide dismutase
Abstract in English
Oxidative stress, caused by increased levels of reactive oxygen species, can lead to damage in all cellular components. Several enzymes, as subunit of alkyl hydroperoxide reductase (AhpC and AhpF) and superoxide dismutases (SOD), are responsible for removing ROS. Mechanisms of gene expression of C. crescentus for genes ahpC, sodA, sodB and sodC, were evaluated with transcription fusions with the lacZ reporter gene were constructed, allowing the quantification of expression by b-galactosidase activity assays, furthermore we analyze of the gene expression by qRT-PCR assay. The cultures were grown in PYE and M2 media, and gene expression was evaluated in the presence of hydrogen peroxide (H2O2), tert-butyl hydroperoxide (tBOOH), paraquat, menadione, pyrogallol, FeSO4 or DPPi. In C. crescentus ahpC is induced by peroxides and is under the control of OxyR. The SODs are mainly induced by superoxide, sodB and sodC are induction in stationary phase and are possibly under the control of sigma J, while the sodA gene is regulated by sigma F and sigma J.
 
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Publishing Date
2014-06-28
 
WARNING: The material described below relates to works resulting from this thesis or dissertation. The contents of these works are the author's responsibility.
  • PREVIATO, M., and MARQUES, MV. Regulation of genes involved in oxidative stress. In 58o Congresso Brasileiro de Genética, Foz do Iguaçu, 2012. Resumos do 58o Congresso Brasileiro de Genética., 2012. Abstract.
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