PichiaPink-3 Chemically Competent Cell

PichiaPink-3 Chemically Competent Cell

PichiaPink-3 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 the yeast culture strains by prokaryotes like E. coli, antibiotics such as Ampicillin and Kanamycin are often added to the culture medium to inhibit bacterial contamination and growth.The optimal growth temperature for PichiaPink-3 yeast is between 24-30°C, and temperatures exceeding 32°C can be catastrophic for protein expression within the cells, potentially leading to cell death. The ADE2 gene encodes phosphoribosylaminoimidazole carboxylase, which catalyzes the sixth step 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 in the vacuoles of yeast cells, causing the cells to appear pink. The Pink3 yeast strain is an ade2 auxotrophic strain, in which the ade2 gene has been deleted, and part of the ade2 promoter is also deleted. Therefore, the strain cannot grow in media lacking adenine and grows slowly in rich media. By transforming a plasmid containing the ade2 gene into this strain, the strain acquires the ability to grow in adenine-deficient media, and its color changes from pink, due to the absence of the ade2 gene, to white, thus indirectly indicating the successful transformation of the cloned strains with the plasmid. PichiaPink-3 is a prb1 gene knockout yeast strain. The prb1 gene encodes protease B, a serine protease found in the vacuoles of the cell, and belongs to the Bacillus cereus protease family. The absence of intracellular protease B can protect the expressed products from degradation and promote an increase in expression levels.

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