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

被引:47
作者
Ahn, Jungoh
Hong, Jiyeon
Lee, Hyeokweon [1 ]
Park, Myongsoo
Lee, Eungyo
Kim, Chunsuk
Choi, Euisung
Jung, Joonki
Lee, Hongweon
机构
[1] Korean Res Inst Biosci & Biotechnol, Biotechnol Proc Engn Dept, Taejon 305600, South Korea
[2] Korean Res Inst Biosci & Biotechnol, Microbial Lab, Taejon 305600, South Korea
[3] Korean Res Inst Biosci & Biotechnol, Biomed Process Dev Dept, Taejon 305600, South Korea
关键词
Pichia pastoris; translation elongation factor; constitutive promoter;
D O I
10.1007/s00253-006-0698-6
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:601 / 608
页数:8
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