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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2010.tde-31082010-081726
Document
Author
Full name
Tatiane Marques Porangaba de Oliveira
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2010
Supervisor
Committee
Sircili, Marcelo Palma (President)
Carvalho, Joao Carlos Monteiro de
Leão, Sylvia Luisa Pincherle Cardoso
Title in Portuguese
Clonagem e expressão de β-ficocianina em amostras de Escherichia coli.
Keywords in Portuguese
Arthrospira platensis
Escherichia coli
Apoptose
Biotecnologia
Clonagem
Expressão gênica
Fibroblastos
Ficocianina
Abstract in Portuguese
C-ficocianina (C-PC) é um pigmento solúvel em água e está presente em Arthrospira platensis. Este pigmento é constituído por duas subunidades, α e β, com massas moleculares de 16 e 17 kDa, respectivamente. Recentemente foi demonstrado que a subunidade β da C-PC de Anabaena tem atividade antitumoral. Os principais objetivos deste estudo foram clonar e expressar a subunidade β de C-PC de A. platensis em amostras de Escherichia coli e verificar a capacidade da proteína recombinante e da proteína C-PC em induzir ou não apoptose em células tumorais (HEp-2) e não tumorais (fibroblastos). As células foram tratadas com 100 µg das proteínas recombinante e C-PC por 6 h. Após tratamento, as células foram coradas com azul de toluidina (Metódo: Concentração Crítica de Eletrólitos - CCE). As células em apoptose foram detectadas pela coloração e por alterações morfológicas. Os resultados obtidos demonstram que ambas as proteínas, β-PC recombinante e C-PC, são capazes de induzir apotose em células HEp-2 e não induzi-la em células fibroblásticas.
Title in English
Cloning and expression of β-phycocyanin in Escherichia coli strains.
Keywords in English
Arthrospira platensis
Escherichia coli
Apoptosis
Biotechnology
Cloning
Fibroblast
Gene expression
Phycocyanin
Abstract in English
C-phycocyanin (C-PC) is a water soluble pigment and is present in Arthrospira platensis. It is consisted of two subunits, α and β, with molecular masses of 16 and 17 kDa, respectively. Recently it was shown that the β subunit of C-PC Anabaena has antitumor activity. The main objectives of this study were cloning and expression the β subunit of C-PC (β-PC) of A. platensis in Escherichia coli and check the ability of the C-PC and recombinant proteins in apoptotic induction in cancer (HEp-2) and non-cancer (fibroblasts) cells. The fibroblasts and HEp-2 cells were treated with 100 µg of recombinant and C-PC, proteins respectively for 6 h. After treatment, the cells were stained with toluidine blue (Critical Electrolyte Concentration Method - CEC). Apoptosis cells were detected by staining and morphological changes. The results show that the recombinant β-PC and C-PC proteins are able to induce apotosis in HEp-2 cells and dont induce it in fibroblasts cells.
 
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Publishing Date
2010-09-16
 
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