Translation elongation factor 1-α gene from Pichia pastoris: Molecular cloning, sequence, and use of its promoter

Abstract

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.