Transformation of the acidophilic heterotroph Acidiphilium facilis by electroporation.
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.
Inagaki
K
K
Division of Bioresources Science, Faculty of Agriculture, Okayama University, Japan.
Tomono
J
J
Kishimoto
N
N
Tano
T
T
Tanaka
H
H
D018274:Electroporation; D005822:Genetic Vectors; D010957:Plasmids; D015183:Restriction Mapping; D013855:Thiobacillus / Q000201:enzymology / Q000235:genetics*; D014169:Transformation, Bacterial; D001618:beta-Lactamases / Q000235:genetics*
D001618:beta-Lactamases