PichiaPink-1 Chemically Competent Cell

PichiaPink-1 Chemically Competent Cell

PichiaPink-1 is a pink strain of Pichia pastoris, belonging to eukaryotic cells. Common antibiotics targeting prokaryotes, such as kanamycin and ampicillin, are ineffective against yeast. Therefore, to prevent contamination of yeast cultures by prokaryotes like E. coli, it is common to add some ampicillin and kanamycin antibiotics to the culture medium to inhibit bacterial contamination and growth.The optimal growth temperature for PichiaPink-1 yeast is 24-30°C, and temperatures exceeding 32°C can have devastating effects on protein expression within the cells, potentially leading to cell death. The ADE2 gene encodes for a phosphoribosyl formylglycinamidine cyclase gene, which catalyzes the sixth reaction in purine nucleotide synthesis. In Saccharomyces cerevisiae, Pichia pastoris, or other yeasts, mutations or deletions of this gene can lead to the accumulation of purine precursors within the vacuoles of yeast cells, causing the cells to appear pink. The Pink1 yeast strain is an ade2 auxotrophic strain, where the ade2 gene is deleted, and part of the ade2 promoter is also missing, so the strain cannot grow in media lacking adenine and grows slowly in rich media. After transforming an expression plasmid containing the ade2 gene into this strain, it allows the strain to grow in media lacking adenine, and the strain's color changes from pink (due to the absence of the ade2 gene) back to white. This indirectly indicates the successful transformation of the plasmid into those cloned strains.

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