Transformation of Paramecium tetraurelia by electroporation or particle bombardment.
J Eukaryot Microbiol
; 46(1): 56-65, 1999.
Article
em En
| MEDLINE
| ID: mdl-10188261
ABSTRACT
Methods for mass transformation of Paramecium tetraurelia were established using plasmids bearing neomycin-resistance or calmodulin gene fragments. Phenotypic and molecular analyses showed that, although variable, up to 5% transformation can be achieved by electroporation. Concentrations of divalent cations Ca2+ and Mg2+ in the electroporation medium were crucial for efficient transformation. Strong neomycin-resistance transformation using bioballistic particle bombardment with gold particles was observed. For both methods, hybridization to transformant DNA revealed plasmid signals consistent with macronuclear transformation and correlated with transformed phenotypes. Complementation of a known calmodulin gene mutation was also achieved by mass transformation. Possible sources of variation and the general utility of these methods are discussed.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Transformação Genética
/
Calmodulina
/
Eletroporação
/
Canamicina Quinase
Limite:
Animals
Idioma:
En
Revista:
J Eukaryot Microbiol
Assunto da revista:
MICROBIOLOGIA
/
PARASITOLOGIA
Ano de publicação:
1999
Tipo de documento:
Article
País de afiliação:
Estados Unidos