Summary
An electroporation method using a Bio-Rad Gene Pulser has been optimized for introducing heterologous DNA into Kluyveromyces lactis yeasts. The plasmid pCR1, derived from a native Kluyveromyces plasmid, was used to transform K. lactis. This plasmid produces a wheat α-amylase and contains both the biosynthetic marker URAA and G418 resistance genes. Transformation was optimal at 4500 V/cm, 25 μF, and ∞ Ω with 0.2 μg plasmid DNA. Transformation efficiencies in the range 104–105 transformants/107 cells/μg DNA were obtained.
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Russell, C., Jarvis, A., Yu, PL. et al. Optimization of an electroporation procedure for Kluyveromyces lactis transformation. Biotechnol Tech 7, 417–422 (1993). https://doi.org/10.1007/BF00151876
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DOI: https://doi.org/10.1007/BF00151876