产甘油假丝酵母HOG1 MAPK同源基因CgHOG1的克隆及特征分析

Cloning and characterization of an HOG1 MAPK homologous gene CgHOG1 from Candida glycerinogenes

摘要:

摘要:【目的】获得产甘油假丝酵母(Candida glycerinogenes)耐高渗和过量合成甘油的关键调控基因—丝裂原活化蛋白激酶基因(CgHOG1)，并考察其渗透压调节功能。【方法】运用简并PCR 结合Self-Formed Adaptor PCR技术从产甘油假丝酵母基因组中克隆CgHOG1基因并进行生物信息学相关分析，将CgHOG1基因在酿酒酵母(Saccharomyces cerevisiae W303-1A)hog1Δ缺失突变株中互补表达，考察菌株耐渗透压能力变化。【结果】所获得CgHOG1基因全长1164 bp，编码387个氨基酸序列(GenBank No. KC480066);氨基酸序列与来源于Ogataea parapolymorpha的Hog1p同源性最高，为86%;该基因在酿酒酵母hog1Δ缺失突变株中异源表达能够显著提高菌株的抗盐耐高渗和甘油合成能力。【结论】本文所获得的基因CgHOG1是一个具有耐高渗和过量合成甘油调控功能的新基因，研究结果为产甘油假丝酵母超高渗应答机制的研究及抗盐耐旱作物改造提供了新的基因。

Abstract:

Abstract:［Objective］This study was aimed to obtain a key gene of osmo-adaptation and glycerol synthesis regulation in Candida glycerinogenes，namely mitogen-activated protein kinase HOG1 gene (CgHOG1)，and to verify its function of osmo-regulation.［Methods］The gene CgHOG1 was amplified by Degenerate PCR and Self-Formed Adaptor PCR from the C. glycerinogenes genome and the bioinformatic analysis of CgHOG1 gene was conducted. The CgHOG1 gene was transformed in Saccharomyces cerevisiae hog1Δ null mutant and its salt tolerance characteristics was investigated.［Results］The gene CgHOG1 encoded a protein of 387 amino acids with an open reading frame of 1164 bp and the amino acid sequence showed 86% identity to Hog1p of Ogataea parapolymorpha． Heterologous expression of CgHOG1 gene in S.cerevisiae W303 hog1Δ null mutant showed an increase in salt tolerance and glycerol production compared to S.cerevisiae W303 hog1Δ null mutant.［Conclusion］The CgHOG1 obtained in this study is a novel HOG1 gene from C. glycerinogenes，which plays an essential role in the yeast hyperosmotic response and glycerol synthesis. We supplied a new gene for the osmo-adaptation mechanism in C. glycerinogenes and molecular modification of the salt-tolerant and droughtresistant crops.