干旱区生态环境知识资源中心(Arid): 干燥和复吸水条件下发菜过氧化物氧还蛋白差异表达与基因克隆

Arid

	干燥和复吸水条件下发菜过氧化物氧还蛋白差异表达与基因克隆
其他题名	Differential Expression and Gene Cloning of Peroxiredoxin from Nostoc flagelliforme Subjected to Dehydration and Rehydration
	梁文裕 1; 焦广飞 2; 周有文 1; 游向荣 3; 张亚萍 1; 陈伟 2
来源期刊	应用与环境生物学报
ISSN	1006-687X
出版年	2011
卷号	17 期号:5 页码:666-672
中文摘要	发菜(Nostoc flagelliforme)是一种陆生固氮蓝藻，为探讨其耐旱的分子机理及对极端干旱环境的适应和保护机制，运用双向电泳技术(2-DE)、凝胶图像分析、MALDI-TOF-TOF/MS质谱鉴定和数据库检索，分析发菜过氧化物氧还蛋白(Peroxiredoxin)在持续干燥48h和复吸水4 h后的差异表达水平，根据鉴定的过氧化物氧还蛋白已知氨基酸序列设计简并性引物克隆该基因，分析基因和氨基酸序列的同源性及对蛋白质二级结构进行预测，并研究其原核表达.结果表明，发菜过氧化物氧还蛋白在复吸水后的表达量明显高于干燥状态下的表达量。根据简并性引物克隆获得长度为639 bp的过氧化物氧还蛋白基因，GenBank登陆号为HM854286.序列比较分析显示该基因具有较高的保守性，蛋白质二级结构主要由alpha螺旋、beta折叠和随机卷曲构成.将过氧化物氧还蛋白基因在大肠杆菌中表达，获得符合预期的外源重组蛋白(26.5*10~3)，经Western blotting验证，该外源蛋白为过氧化物氧还蛋白。
英文摘要	Nostoc flagelliforme is a terrestrial nitrogen-fixing cyanobacteria with strong drought ecological adaptability.In this research, 2-DE, gel image analysis and MALDI-TOF-TOF/MS were applied to compare and identify the peroxiredoxin expression level from N.flagelliforme subjected to dehydration and rehydration.Degeneracy primer was designed based on the identified amino acid sequences in order to clone peroxiredoxin gene, to analyze the gene and amino acid sequences, to predict the secondary structure of peroxiredoxin, and to study the prokaryotic expression of the peroxiredoxin gene.The results indicated that peroxiredoxin expression level of N.flagelliforme subjected to rehydration was higher than that to desiccation. Peroxiredoxin gene was cloned and a full length of 639 bp DNA was obtained(GenBank access number: HM854286). Homology analysis showed that the N.flagelliforme peroxiredoxin gene has high consensus regions.The secondary structure of peroxiredoxin was made up of alphahelix, betastrands and random coil.Peroxiredoxin gene was expressed in E.coli, and a M_r 26.5*10~3 heterologous protein was obtained.It was confirmed by Western blotting that the protein was peroxiredoxin.The study laid a foundation for in depth research in terms of the drought-resistant molecular mechanism and protection mechanism of N. flagelliforme in extreme arid environment.
中文关键词	发菜 ; 过氧化物氧还蛋白 ; 双向电泳 ; 差异表达 ; 基因克隆 ; 原核表达 ; 耐旱
英文关键词	Nostoc flagelliforme peroxiredoxin 2-DE differential expression gene clone prokaryotic expression drought-resistance
语种	中文
国家	中国
收录类别	CSCD
WOS类目	PLANT SCIENCES
WOS研究方向	Plant Sciences
CSCD记录号	CSCD:4347030
来源机构	宁夏大学
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/227355
作者单位	1.宁夏大学生命科学学院, 银川, 宁夏 750021, 中国; 2.福建农林大学生命科学学院, 福州, 福建 350002, 中国; 3.广西农业科学院农产品加工研究所, 南宁, 广西 530007, 中国
推荐引用方式 GB/T 7714	梁文裕,焦广飞,周有文,等. 干燥和复吸水条件下发菜过氧化物氧还蛋白差异表达与基因克隆[J]. 宁夏大学,2011,17(5):666-672.
APA	梁文裕,焦广飞,周有文,游向荣,张亚萍,&陈伟.(2011).干燥和复吸水条件下发菜过氧化物氧还蛋白差异表达与基因克隆.应用与环境生物学报,17(5),666-672.
MLA	梁文裕,et al."干燥和复吸水条件下发菜过氧化物氧还蛋白差异表达与基因克隆".应用与环境生物学报 17.5(2011):666-672.