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Doctoral Thesis
DOI
10.11606/T.42.2001.tde-01112001-123050
Document
Author
Full name
João Marcelo Pereira Alves
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2001
Supervisor
Committee
Affonso, Maria Heloiza Trebilcock (President)
Briones, Marcelo Ribeiro da Silva
Camargo, Erney Felicio Plessmann de
Matioli, Flora Maria de Campos Fernandes
Shaw, Jeffrey Jon
Title in Portuguese
Caracterização e filogenia moleculares de Acanthamoeba.
Keywords in Portuguese
Acanthamoeba (de preferência em itálico)
Filogenia
Identificação de organismos
RNA ribossômico
Sequenciamento de DNA
Abstract in Portuguese
Neste trabalho foram caracterizadas molecularmente e inferidas as relações filogenéticas de 14 isolados brasileiros de Acanthamoeba, provenientes de casos de ceratite, e 8 isolados da ATCC (4 de ceratite e 4 ambientais). Foram utilizados inicialmente os métodos de RAPD, RFLP de DNA genômico total e RFLP do SSU rDNA. Apesar de revelar a alta variabilidade genética em Acanthamoeba, estes métodos permitiram estabelecer grupos bem definidos de isolados mais similares geneticamente. O seqüenciamento do SSU rDNA permitiu a inferência da filogenia entre os isolados utilizados nesse estudo em relação àqueles presentes na literatura, que estão distribuídos em doze tipos de seqüência deste gene. Dentre os 17 isolados de ceratite presentes em nosso estudo, 16 apresentaram SSU rDNA tipo T4 (anteriormente já fortemente correlacionado à ceratite) e um deles constitui um novo tipo de seqüência. Dois dos 4 isolados ATCC (ambientais) cujas seqüências ainda não haviam sido determinadas também apresentaram novos tipos de SSU rDNA, enquanto outros 2 apresentaram o tipo T4.
Title in English
Molecular characterization and phylogeny of Acanthamoeba.
Keywords in English
Acanthamoeba
DNA sequencing
Organism identification
Phylogeny
ribosomal RNA
Abstract in English
In this work we performed the molecular phylogeny and characterization of 22 Acanthamoeba isolates, 14 Brazilian keratitis isolates and 8 from ATCC, 4 keratitis and 4 environmental isolates. In spite of the extensive genetic variability disclosed by RAPD, total genomic DNA RFLP and SSU rDNA RFLP techniques, these methods enabled us to group some isolates in well defined clusters of genetically more related organisms. Sequencing of SSU rDNA allowed inference of the phylogeny of our isolates with those present in the literature, which are distributed through 12 sequence types of this gene. Among the 17 keratitis isolates of our study, 16 presented SSU rDNA of type T4 (previously found to be strongly correlated to keratitis), and one was assigned to a new sequence type. Of the 4 isolates from ATCC whose sequences were previously undetermined, the two environmental isolates also constituted new sequence types, while the two keratitis isolates were assigned to type T4.
 
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tde.pdf (3.26 Mbytes)
Publishing Date
2003-10-15
 
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