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Detection of genetic polymorphism among and within Echinococcus granulosus strains by heteroduplex analysis of a microsatellite from the U1 snRNA genes
P.A. Roratto1, M.L. Bartholomei-Santos1, A.M. Gutierrez2,
L. Kamenetzky2, M.C. Rosenzvit2 and A. Zaha3
1Departamento de Biologia, CCNE, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil
2Departamento de Parasitologia, Instituto Nacional de Enfermedades Infecciosas,
ANLIS ‘Dr. Carlos G. Malbrán’, Buenos Aires, Argentina
3Centro de Biotecnologia e Departamento de Biologia Molecular e Biotecnologia,
Instituto de Biociências, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil
Corresponding author: M.L. Bartholomei-Santos
E-mail: marlise@smail.ufsm.br
Genet. Mol. Res. 5 (3): 542-552 (2006)
Received September 21, 2005
Accepted August 1, 2006
Published September 14, 2006

ABSTRACT. Polymerase chain reaction of a pentanucleotide microsatellite in the U1 snRNA gene complex generated a multiple band pattern due to the priming of paralogous sequences. Denaturation and slow renaturation of polymerase chain reaction products allow the formation of heteroduplex DNA that can be detected by its differential mobility in polyacrylamide gel electrophoresis. Heteroduplex analysis was used to determine if the U1 snRNA microsatellite could be a useful genetic marker in Echinococcus granulosus. A U1 snRNA microsatellite fragment from E. granulosus was isolated and characterized by Southern blot and sequencing. Four E. granulosus strains were analyzed: sheep, Tasmanian sheep, cattle, and camel strains. The former two showed polymorphism and shared three of the six patterns found for sheep strain. The cattle strain displayed two patterns, and the camel strain was monomorphic. The electrophoretic profiles were used for statistical analysis in order to determine genetic distance and the relationship among strains. Heteroduplex analysis can be helpful in genotyping E. granulosus strains and is useful in detecting polymorphism within strains.

Key words: Echinococcus granulosus, Microsatellite markers, Heteroduplex DNA, U1 snRNA gene

 

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