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-alpha 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.

Expression of Zea mays transglutaminase in Pichia pastoris under different promoters and its impact on properties of acidified milk protein concentrate gel

“…Its gene was shown to have strong promoter activity in other organisms. The study demonstrated that P TEF1 has a similar capability to produce foreign protein as those produced by P GAP . However, in this study, TGZ expression level of P TEF1 ‐ tgz was lower than that of P FLD1 ‐ tgz , so the recombinant strain under the control of P FLD1 (P FLD1 ‐ tgz ) was selected for subsequent TGZ production.…”

“…As an inducible promoter, it can be independently induced either by methanol as a carbon source or by methylamine as a nitrogen source . Another promoter, the constitutive translation elongation factor 1‐α promoter (P TEF1 ), has attracted research attention by demonstrating a tighter growth‐associated expression mode and a similar or even greater yield on recombinant protein than glyceraldehyde‐3‐phosphate dehydrogenase promoter (P GAP ), a constitutive promoter that provides a comparable expression level to P AOX1 . P FLD1 and P TEF1 can eliminate the disadvantage related to the use of methanol during the expression of heterologous protein under the control of P AOX1 , thus becoming promising alternative promoters for the expression of foreign protein induced by methylamine or glycerol.…”

Promoter library designed for fine-tuned gene expression in Pichia pastoris

“…Although several promoters, either constitutive or inducible, are available for this yeast [P AOX1 (8), P AOX2 (9), P FLD1 (10), P DAS (8), P PEX8 (P PER3 ) (11), P YPT1 (12), P GAP (13), P TEF1 (14), P PGK1 (15), P ICL1 (16)], the promoter of the alcohol oxidase I gene ( AOX1 ) has been employed in most studies and applications (1). AOX1 is the predominantly expressed form of two alcohol oxidases ( AOX1 and AOX2 ) in the P. pastoris cell.…”

Manipulating Natural Product Biosynthetic Pathways via DNA Assembler

“…Compared to the control promoter, much stronger activities of gapdh p (SG) and rpsL p (SG) should be observed. The strong activities of gapdh p and the promoters of various translation elongation factors have also been observed in many other microorganisms, such as fungi, bacteria, micro-algae, and protozoa (Ahmad et al, 2012; Ahn et al, 2007; Hong et al, 2010; Jia et al, 2012; Krasny et al, 2000; Shao et al, 2009; Suarez et al, 2006). …”