Open AccessTransfection and transient expression of chloramphenicol acetyltransferase gene in the protozoan parasite Entamoeba histolytica.
Author(s) -
Rose Nickel,
Egbert Tannich
Publication year - 1994
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.91.15.7095
Subject(s) - chloramphenicol acetyltransferase , entamoeba histolytica , transfection , electroporation , microbiology and biotechnology , biology , gene , plasmid , gene expression , reporter gene , biochemistry
Hybrid plasmids were constructed and used for successful transfection and transient expression of the chloramphenicol acetyltransferase (CAT) gene in the protozoan parasite Entamoeba histolytica. Transfection was performed by electroporation of the amebae in a potassium phosphate-based buffer under conditions of 3000 V/cm and 25 microF, resulting in a time constant of 0.4 ms. Expression of CAT activity was achieved with constructs in which the CAT coding region was flanked by untranslated upstream and downstream sequences of E. histolytica genes. Highest activity was detected after culturing transfected cells for 48 hr. Activity was found to be proportional to the amount of DNA transfected.