糙皮侧耳乙醛脱氢酶基因PoALDH1的克隆与非生物胁迫下的表达分析

发布时间：2019-01-06 07:00

【摘要】：本研究从糙皮侧耳中克隆了乙醛脱氢酶基因PoALDH1(Gen Bank登录号为KT026035)和其全长c DNA序列。长为2 016bp的PoALDH1序列编码478个氨基酸,分子量约为52k Da,氨基酸序列中含有乙醛脱氢酶保守的谷氨酸活性位点和半胱氨酸残基活性位点。PoALDH1基因在大肠杆菌中表达后显示,重组菌株的乙醛脱氢酶比活力为0.58U/mg,且重组菌株的乙醛耐受性显著高于对照菌株。实时荧光定量PCR(quantitative real-time PCR)分析PoALDH1基因在糙皮侧耳不同发育时期的表达结果显示,PoALDH1基因在原基期的表达量约为双核菌丝期的5倍。PoALDH1基因在原基发育起始阶段经光照及温差刺激后均上调表达。当糙皮侧耳菌丝暴露在不同浓度的氯化钠及甘露醇的条件下时,菌丝生长均受到抑制且PoALDH1基因的表达量均高于对照。研究结果将为进一步从分子水平揭示糙皮侧耳的抗逆机制以及食用菌的发育奠定一定基础。
[Abstract]:In this study, acetaldehyde dehydrogenase gene PoALDH1 (Gen Bank accession number was KT026035 and its full-length c DNA sequence was cloned from Pleurotus strigella. The PoALDH1 sequence with a length of 2 016bp encodes 478amino acids. The molecular weight of the amino acid sequence is about 52k Da, which contains the conserved glutamate active site and cysteine residue active site of acetaldehyde dehydrogenase. The PoALDH1 gene is expressed in E. coli. The specific activity of acetaldehyde dehydrogenase of the recombinant strain was 0.58 U / mg, and the acetaldehyde tolerance of the recombinant strain was significantly higher than that of the control strain. The expression of PoALDH1 gene in different developmental stages of Pleurotus strigella was analyzed by real-time fluorescence quantitative PCR (quantitative real-time PCR. The expression of PoALDH1 gene in the primordial stage was about 5 times higher than that in the binaryotic mycelium stage, and the expression of PoALDH1 gene was up-regulated during the initial stage of the primordium development after being stimulated by light and temperature difference. When the hyphae of Pleurotus strigella was exposed to different concentrations of sodium chloride and mannitol, the hyphal growth was inhibited and the expression of PoALDH1 gene was higher than that of the control. The results will lay a foundation for further understanding the stress resistance mechanism of Pleurotus strigella and the development of edible fungi at molecular level.
【作者单位】：河南农业大学生命科学学院;
【基金】：现代农业产业技术体系建设专项(CARS-24) 河南省重点科技攻关项目(162102110091)~~
【分类号】：S646

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