Translation elongation factor 1-α gene from Pichia pastoris: molecular cloning, sequence, and use of its promoter
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The gene encoding translation elongation factor 1-α from the yeast Pichia pastoris was cloned. The gene revealed an open reading frame of 1,380 bp with the potential to encode a polypeptide of 459 amino acids with a calculated mass of 50.1 kDa. The potential of the promoter (P TEF1 ) in P. pastoris was investigated with comparison to the glyceraldehyde-3-phosphate dehydrogenase promoter (P GAP ) by using a bacterial lipase gene as a reporter gene. P TEF1 demonstrated a tighter growth-associated expression mode, improved functioning in the presence of high glucose concentrations, and promoter activities that yielded recombinant protein at levels similar to or in one case greater than P GAP . The sequence of the gene was deposited in GenBank under accession no. EF014948.
KeywordsPichia pastoris Translation elongation factor Constitutive promoter
We are grateful to Philekorea Technology (Korea) for technical assistance with the real-time quantitative PCR method.
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