Transformation of the Acidophilic HeterotrophAcidiphilium facilisby Electroporation

Abstract

We constructed a cloning vector for use in the acidophilic heterotroph Acidiphilium facilis. The vector pAH101 (8.8 kb) was constructed from a 6.1 kb restriction fragment of the Acidiphilium plasmid pAH1 and a pUC19 carrying a beta-lactamase gene. The antibiotic resistance gene was efficiently expressed in A. facilis. Several factors which influenced the transformation efficiency were optimized, resulting in a transformation efficiency of up to 3 x 10(3) transformants per microgram of plasmid DNA at a field strength of 10 kV/cm with a 7.0 ms pulse.