Application Focus - Genetic Engineering of Brewer's Yeast with Electroporation
By Michelle M. Ng, Ph. D.
In honor of National Beer Day, April 7th, the focus of this blog post is how to use electroporation for genetic engineering of brewing yeast. First, we will provide the ideal electroporation parameters for Saccharomyces cerevisiae using either ECM 630 or Gemini X2 electroporators. Then, we will highlight an example from the primary literature where this electroporation method was used with brewing yeast Saccharomyces cerevisiae ssp. carlsbergensis to modify the amount of ethanol in beer.
- Grow Saccharomyces cerevisiae up to log phase in YPD media
- Cetrifuge and wash pelleted cells several times to remove cell culture media and suspend cells in electroporation buffer at desired density of 1.6 x 109 cells/ml
- Electroporate cells in 2 mm gap cuvettes or 2 mm gap High Throughput electroporation plates with the following conditions:
- Temperature: 4°C
- Voltage: 2500 V
- Capacitance: 25 uF
- Resistance: 100 to 200 Ω
- Number of Pulses: 1
- Desired Electrical Field Strength: 12.5 kV/cm
- Desired Time Constant: 3 to 4.5 ms
1. Nevoigt, E., et al. Engineering of brewing yeast to reduce the content of ethanol in beer. FEMS Yeast Research 2002; 2: 225–232.
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