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Master's Dissertation
DOI
10.11606/D.87.2012.tde-05022013-082130
Document
Author
Full name
Mayra Akemi Kuroki
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Sluys, Marie Anne van (President)
Hotta, Carlos Takeshi
Maldonado, Gabriel Padilla
Title in Portuguese
Desenvolvimento de uma estratégia de clonagem customizada de regiões promotoras do genoma da cana-de-açúcar.
Keywords in Portuguese
Cana-de-açúcar
Clonagem
Genomas
Monocotiledôneas
Protoplastos
Abstract in Portuguese
O objetivo deste trabalho foi desenvolver uma metodologia para identificação de regiões promotoras funcionais a partir de segmentos de um genoma qualquer. O genoma da cana-de-açúcar foi escolhido para o desenvolvimento desta estratégia na qual envolve a obtenção de fragmentos de DNA os quais foram clonados em vetores de expressão. A triagem destes fragmentos foi realizada através de biobalística e resultou no isolamento de quatro clones. Um ensaio de transformação permanente em arroz com três clones gerou 12 plantas. Foi detectada expressão do marcador GUS em calos, folhas e raízes, comprovando sua funcionalidade. Desta maneira, o presente trabalho permitiu estabelecer uma metodologia de recuperação de sequências regulatórias funcionais com ampla possibilidade de serem explorados biotecnologicamente.
Title in English
Customized promoter cloning strategy.
Keywords in English
Cloning
Genomes
Monocotyledonous
Protoplasts
Sugarcane
Abstract in English
The aim of this work is develop a strategy to identify functional promoter regions from any genome. The modern sugarcane genome was chosen as a model for the development of this strategy that involves the generation of fragments of DNA and cloning them into expression vectors. These fragments were then screened by a transient expression assay using biolistic particle delivery resulting in the isolation of four clones. Three clones were permanently transformed in rice, and 12 plants were obtained. GUS expression was detected in the callus, leaves and roots of the rice plants thus confirming the functionality of sequences in these clones. The present work has established a strategy to identify and extract functional regulatory sequences containing functional regulatory regions which show great potential of being useful in both the biotechnology field and in the field of basic science.
 
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Publishing Date
2013-02-22
 
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