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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2011.tde-30052012-084230
Document
Author
Full name
Yeimy Paola Galindo Rozo
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Silva, Luiziana Ferreira da (President)
Lopes, Mateus Schreiner Garcez
Marques, Marilis do Valle
Title in Portuguese
Bioprospecção de genes relacionados à biossíntese de polímeros biodegradáveis a partir de uma biblioteca metagenômica de solo de Mata Atlântica.
Keywords in Portuguese
Biopolímeros
Biotecnologia
Gênese do solo
Genomas
Polihidroxialcanoatos
Polímeros (Química Orgânica)
Abstract in Portuguese
O presente estudo teve como objetivo realizar a triagem de PHA sintases numa biblioteca metagenômica de solo de Mata Atlântica, empregando duas técnicas de busca: o método de detecção fenotípica e a técnica de PCR. Resultados positivos foram obtidos com este último, empregando iniciadores descritos na literatura e iniciadores descritos neste trabalho. Em 10.67% dos clones da biblioteca foram obtidos amplicons, dos quais 7 foram seqüenciados, apresentando similaridade para os genes phaC dos tipos II e IV. Adicionalmente, 67 clones positivos para a classe III foram obtidos e 4 destes foram seqüenciados. Duas das seqüências obtidas mostraram alta similaridade com o gene codificador da enzima glutamina sintetase tipo I, outro deles para a proteína hipotética conservada pertencente à família de enzimas oxidoredutases e a outra para o componente D do gene hidrogenase-4. A partir da análise dos resultados, iniciadores mais específicos são propostos. Assim, a técnica de PCR foi mais eficiente na detecção de genes da biossíntese de PHA na biblioteca metagenômica estudada.
Title in English
Bioprospecting in a metagenomic library from Atlantic forest soil for genes involved on the biosynthesis of biodegradable polymers.
Keywords in English
Biopolymers
Biotechnology
Genomes
Polyhydroxyalkanoates
Polymers (Organic Chemistry)
Soil genesis
Abstract in English
To perform a PHA synthase screening in a metagenomic library from Atlantic forest soil two search methods were applied: phenotypic detection and PCR. Positive results with PCR were obtained by using primers described in the literature and proposed in this study. Amplicons were obtained in 10.67% of the library, 7 of them were sequenced showing similarity with class II and IV phaC genes. In addition, 67 positive clones for class III were obtained and 4 of them were sequenced. Two of these sequences showed high similarity to the glutamine synthase gene type I, the third one showed similarity to the conserved hypothetical protein of the reductase family, and the forth presented similarity to the component D of the hidrogenase-4. According to the results, more specific primers are suggested. Therefore, PCR was more efficient in the detection of PHA biosynthesis genes in the studied metagenomic library.
 
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Publishing Date
2012-06-27
 
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