Translation elongation factor 1-alpha gene from Pichia pastoris: molecular cloning, sequence, and use of its promoter.
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.
Ahn
Jungoh
J
Biotechnology Process Engineering Department, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 305-600, South Korea.
Hong
Jiyeon
J
Lee
Hyeokweon
H
Park
Myongsoo
M
Lee
Eungyo
E
Kim
Chunsuk
C
Choi
Euisung
E
Jung
Joonki
J
Lee
Hongweon
H
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000235:genetics; D008049:Lipase / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D020648:Peptide Elongation Factor 1 / Q000737:chemistry / Q000235:genetics*; D010843:Pichia / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012440:Saccharomyces / Q000235:genetics; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D005656:Fungal Proteins; D020648:Peptide Elongation Factor 1; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D008049:Lipase