Genetic Transformation of Candida glycerinogenes by REMI and Electroporation

Abstract:

In order to isolate genes related with the osmoadaptation and glycerol metabolism of Candida glycerinogenes, a transformation system based on the dominant selectable marker Zeocin and restriction enzyme-mediated integration (REMI) was established. Effects of seven restriction enzymes on transformation efficiency of C.glycerinogenes were tested. Transformation conditions were optimized in the presence of Hind III. Under the optimal conditions of OD₆₀₀≈1.3, voltage of 1.5 kV, 2.0×10⁹ competent cells/mL, 100 units of Hind III added, the transformation efficiency was up to 129 trnaformants/μg DNA. 58% of transformants were stable on nonselective medium. These results suggest that REMI technique would be beneficial to the genetic transformation of C.glycerinogenes.