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Transformation of Paramecium tetraurelia by Electroporation or Particle Bombardment
Author(s) -
BOILEAU ANDREW J.,
KISSMEHL ROLAND,
KANABROCKI JOSEPH A.,
SAIMI YOSHIRO
Publication year - 1999
Publication title -
journal of eukaryotic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 1066-5234
DOI - 10.1111/j.1550-7408.1999.tb04584.x
Subject(s) - electroporation , transformation (genetics) , paramecium , plasmid , biology , complementation , gene , genetics , microbiology and biotechnology , phenotype
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 Ca 2‐ and Mg 2+ 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.