High-level expression of human tumour suppressor P53 in the methylotrophic yeast: Pichia pastoris.
The human tumour suppressor P53 is a key protein involved in tumour suppression. P53 acts as a "guardian of genome" by regulating many target genes involved in cell cycle regulation, DNA repair and apoptosis. We report the P53 expression by the methylotrophic yeast Pichia pastoris using the methanol inducible AOX1 promoter. We have produced the rP53 in intracellular form as well as secreted using the Saccharomyces cerevisiae alpha-mating factor prepro-leader sequence in two genetic contexts of Pichia, Mut(s) and Mut(+). The intracellular P53 was successfully produced by Mut(s) (KM71) as well as Mut(+) (X33) strains, however, the secreted form was mainly observed in the Mut(s) strain, despite a higher number of p53 copies integrated in the Mut(+) strain. Interestingly, in Mut(s) phenotype, the medium pH influences markedly the rP53 production since it was higher at pH 7 than 6.
Abdelmoula-Souissi
Salma
S
Laboratoire de Génétique Moléculaire des Eucaryotes, Centre Biotechnologie de Sfax, route Sidi Mansour, Sfax, Tunisia.
Rekik
Leila
L
Gargouri
Ali
A
Mokdad-Gargouri
Raja
R
D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis*
D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53