• JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
 
  Bookmark and Share
 
 
Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2012.tde-26072012-142501
Document
Author
Full name
Adam Arai Martens
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Siviero, Fábio (President)
Guimaraes, Margareth de Lara Capurro
Mori, Lyria
Title in Portuguese
Caracterização do gene CDK7 e análise de sua possível atuação nos endociclos de Rhynchosciara americana.
Keywords in Portuguese
Biologia molecular
Ciclo celular
Desenvolvimento animal
Diptera
Sequenciamento genético
Abstract in Portuguese
CDKs são proteínas responsáveis pela ativação e progressão do ciclo celular e também apresentam função na ativação e alongamento na transcrição. Dentre elas, CDK7 atua em ambas as funções, fosforilando as CDKs do ciclo celular e fazendo parte do fator geral de transcrição TFIIH como subunidade catalítica. A partir de uma biblioteca de ESTs de glândula salivar, construído com mRNAs presentes durante o período em que ocorre o início do último ciclo replicativo da politenização e início da amplificação gênica, foram encontradas poucas mensagens relacionadas diretamente com o ciclo celular, correspondendo a 3.11% do ESTs. Dentre elas, foram encontradas as mensagens de Cdc2-like e Cdk7; portanto, foi realizada a caracterização do gene Cdk7 e a análise de sua atuação durante o desenvolvimento larval de Rhynchosciara americana. O gene Cdk7 apresenta 4 éxons, mais que em vertebrados. A sequência completa do mRNA foi obtida a partir de RACE, apresentando 1230 bases e uma ORF de 1020 bases. Perfis de expressão foram determinados por RT-PCR e Western blots. Modificações pós-traducionais foram analisadas por imunoblots em 2D. Seu perfil de expressão de mRNA e proteína apresentam variações durante o ciclo celular e entre os tecidos analisados; os imunoblots mostram a presença de uma fosforilação e possíveis modificações na cadeia lateral de alguns aminoácidos. O estudo de proteínas relacionadas ao ciclo celular nesse modelo é importante para um melhor entendimento dos ciclos celulares incomuns presentes em diferentes tecidos de insetos.
Title in English
Cdk7 gene characterisation and analysis of its possible role in the endocycles of Rhynchosciara americana.
Keywords in English
Animal development
Cell cycle
Diptera
Genetic sequencing
Molecular biology
Abstract in English
CDKs are proteins responsible for activation and progression of cell cycle and also work on the activation and elongation of transcription. Among them, CDK7 acts in both functions, phosphorylating cell cycle CDKs and as a part of general transcription factor TFIIH as its catalytic subunit. From an EST library of salivary gland, constructed with mRNAs present during the period when the beginning of the last polyteny replicative cycle occurs, it was found only few messages directly related to cell cycle, corresponding to 3.11% of the ESTs. Among them, it was found Cdc2-like and Cdk7; therefore, it was performed the characterisation of Cdk7 gene and the analysis of its role during the larval development of Rhynchosciara americana. Cdk7 gene presents 4 exons, more than in vertebrates. Complete mRNA sequence was obtained via RACE, presenting 1230 bases and an 1020 bases ORF. Expression profiles were determined by RT-PCR and Western blots. Posttranslational modifications were analysed by 2D immunoblots. Its mRNA and protein expression profiles presented variations during cell cycle and between the studied tissues; immunoblots showed the presence of one phosphorylation and possible modifications on the side chain of some amino acids. The study of proteins related to cell cycle in this model is important for a better understanding of uncommon cell cycles in different insect tissues.
 
WARNING - Viewing this document is conditioned on your acceptance of the following terms of use:
This document is only for private use for research and teaching activities. Reproduction for commercial use is forbidden. This rights cover the whole data about this document as well as its contents. Any uses or copies of this document in whole or in part must include the author's name.
Publishing Date
2012-08-15
 
WARNING: Learn what derived works are clicking here.
All rights of the thesis/dissertation are from the authors
CeTI-SC/STI
Digital Library of Theses and Dissertations of USP. Copyright © 2001-2024. All rights reserved.