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Doctoral Thesis
DOI
https://doi.org/10.11606/T.11.2006.tde-06072006-163915
Document
Author
Full name
Paola Alejandra Fiorani Celedon
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2006
Supervisor
Committee
Labate, Carlos Alberto (President)
Carrer, Helaine
Machado, Marcos Antonio
Novello, Jose Camillo
Pinheiro, José Baldin
Title in Portuguese
Identificação de proteínas da região cambial de Eucalyptus grandis por eletroforese bidimensional e espectrometria de massas
Keywords in Portuguese
cambio (botânica)
espectrometria de massas
eucalipto
proteínas de plantas
Abstract in Portuguese
A proteômica expandiu-se dentro da área biológica a partir da década de 90 com o desenvolvimento de técnicas de ionização branda que permitem analisar macromoléculas por espectrometria de massas (MS) e é uma nova estratégia na busca de genes de interesse e de informações sobre o controle da expressão gênica para a manipulação genética de plantas. Na busca por genes diferencialmente expressos durante o desenvolvimento da madeira, foi estabelecida uma plataforma 2D-LC-MS/MS (eletroforese bidimensional seguida de cromatografia líquida e espectrometria de massas em tandem) e identificadas 72 proteínas da região cambial de Eucalyptus grandis de árvores de 2 anos e 8 meses. A identificação foi feita por homologia com o banco de proteínas SwissProt, com um banco de ESTs específico de Eucalyptus spp. (GENOLYPTUS) e com o NCBI. Os dados de proteínas também foram confrontados com a expressão dos transcritos correspondentes obtidos a partir do mesmo tecido por SAGE (Serial Analysis of Gene Expression). Dentre as proteínas identificadas, a maior parcela (24,1 %) tem função relacionada com resposta a estresse, 20,5 % pertence a metabolismo de energia, 17,8 % são componentes estruturais, 14,3 % participam em metabolismo de macromoléculas e 15,2 % em metabolismos básicos como de nitrogênio, aminoácidos, nucleotídeos, lipídeos e metabolismo secundário.
Title in English
Identification of proteins from the cambial region of Eucalyptus grandis by bidimensional electrophoresis and mass spectrometry
Keywords in English
cambium (botany)
Eucalyptus
mass spectrometry
plant proteins
Abstract in English
Proteomics impacted the biological sciences since the 90´s after the development of soft ionization techniques that allow to analyse macromolecules by mass spectrometry (MS). It became a new strategy to identify genes and obtain information about the molecular control of gene expression, of importance to genetic manipulation of plants. In order to search for genes involved in wood quality and to understand gene expression during wood development, a 2D-LC-MS/MS system (bidimensional electrophoresis followed by liquid cromatography coupled to mass spectrometry in tandem) was stablished. 72 proteins from the cambial region of Eucalyptus grandis trees at the age of 2 years and 8 months were identified. The MS/MS spectra were processed using the SwissProt databank, an EST (Expressed Sequence Tags) bank of Eucalyptus spp. (GENOLYPTUS) and the NCBI. A comparation of gene and protein expression was carried out using a databank constructed in our laboratory using SAGE (Serial Analysis of Gene Expression) obtained from the same cambial tissue. Most identified proteins are related to stress response (24.1 %), 20.5 % participate in energy metabolism, 17.8 % are important to the cell as structural components, 14,3 % are related to macromolecular metabolism and 15,2 % to basic metabolism of nitrogen, amino acids, nucleotides, lipids and secondary metabolism.
 
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PaolaCeledon.pdf (2.12 Mbytes)
Publishing Date
2006-07-12
 
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