巴斯德毕赤酵母翻译延伸因子1-α基因：分子克隆、序列及其启动子的应用

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

作者信息

Ahn Jungoh, Hong Jiyeon, Lee Hyeokweon, Park Myongsoo, Lee Eungyo, Kim Chunsuk, Choi Euisung, Jung Joonki, Lee Hongweon

机构信息

Biotechnology Process Engineering Department, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 305-600, South Korea.

出版信息

Appl Microbiol Biotechnol. 2007 Mar;74(3):601-8. doi: 10.1007/s00253-006-0698-6. Epub 2006 Nov 24.

DOI:10.1007/s00253-006-0698-6

PMID:17124582

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.

摘要

克隆了来自巴斯德毕赤酵母的编码翻译延伸因子1-α的基因。该基因显示出一个1380 bp的开放阅读框，有可能编码一个由459个氨基酸组成的多肽，计算分子量为50.1 kDa。通过使用细菌脂肪酶基因作为报告基因，与甘油醛-3-磷酸脱氢酶启动子（P(GAP)）相比，研究了毕赤酵母中启动子（P(TEF1)）的潜力。P(TEF1)表现出更紧密的生长相关表达模式，在高葡萄糖浓度下功能改善，并且启动子活性产生的重组蛋白水平与P(GAP)相似，在一种情况下高于P(GAP)。该基因序列已存入GenBank，登录号为EF014948。