The most commonly used transformation methods for Saccharomyces cerevisiae are the electrotransformation method and the protoplast method, among which the electrotransformation method is the most convenient, the transformation efficiency is the highest, and it is the easiest to produce multi-copy integration. The protoplast method requires the preparation of protoplasts with the cell wall removed to facilitate the entry of DNA, and then the cells regenerate the cell wall to produce transformants, whereas ZeocinR inhibits the formation of the cell wall, resulting in the failure to produce transformants. Therefore, vectors such as pPICZ series, which use ZeocinR as a selection marker, cannot be transformed by the protoplast method. Lithium acetate was ineffective for Saccharomyces cerevisiae, but effective for Saccharomyces cerevisiae, and only lithium chloride was effective; PEG3350 could shield the toxic effect of high concentration of LiCl.

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