Transformation of Paramecium tetraurelia by electroporation or particle bombardment.
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.
Boileau
A J
AJ
Neuroscience Training Program, University of Wisconsin-Madison 53706, USA. boileau@facstaff.wisc.edu
Kissmehl
R
R
Kanabrocki
J A
JA
Saimi
Y
Y
D000818:Animals; D002147:Calmodulin / Q000235:genetics*; D018274:Electroporation / Q000379:methods*; D019868:Kanamycin Kinase / Q000235:genetics*; D016296:Mutagenesis; D016805:Paramecium tetraurelia / Q000235:genetics; D010641:Phenotype; D014170:Transformation, Genetic
D002147:Calmodulin; D019868:Kanamycin Kinase