植物抗逆蛋白（LEA3）22-氨基酸耐盐结构域在酵母细胞中的鉴定

doi:10.7525/j.issn.1673-5102.2009.01.016

植物研究 ›› 2009, Vol. 29 ›› Issue (1): 74-79.doi: 10.7525/j.issn.1673-5102.2009.01.016

植物抗逆蛋白（LEA3）22-氨基酸耐盐结构域在酵母细胞中的鉴定

刘昀;李冉辉;汪为茂;梁翔宇;郑易之^*

深圳大学生命科学学院，深圳市微生物基因工程重点实验室，深圳　518060

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-01-20 发布日期:2009-01-20
通讯作者: 郑易之
基金资助:

Functional Identification of Plant Stress-resistant Protein(LEA3) and Its 22-mer Motifs in Yeast

LIU Yun;LI Ran-Hui;WANG Wei-Mao;LIANG Xiang-Yu;ZHENG Yi-Zhi*

College of Life Sciences,Key Laboratory of Microorganism and Genetic Engineering of Shenzhen,Shenzhen University,Shenzhen　518060

Received:1900-01-01 Revised:1900-01-01 Online:2009-01-20 Published:2009-01-20
Contact: ZHENG Yi-Zhi
Supported by:

摘要/Abstract

摘要： 大豆PM2蛋白属LEA3 （late embryogenesis abundant 3）蛋白。本文构建了编码PM2全长及含22-氨基酸结构域多肽（PM2、PM2A、PM2B和 PM2C）的酵母表达载体。转化酵母得到四种重组菌。SDS-PAGE电泳和ESI-MS/MS或MALDI-TOF/TOF质谱结果表明，重组菌可表达目标多肽。测定对照菌及重组菌在无胁迫、高盐（1.5 mol·L^-1 NaCl）和高渗透（2 mol·L^-1山梨糖）下的生长曲线。结果表明，在高盐胁迫下，四种重组菌胁迫后的恢复明显好于对照菌。多肽对高盐耐受能力的大小为:PM2C>PM2B≈PM2A≈PM2。证明大豆PM2蛋白的表达可提高酵母耐盐性，且22-氨基酸基序为PM2蛋白的耐盐结构域。结合前文在大肠杆菌中的结果，为“LEA蛋白可以类似机制参与原核和真核生物耐盐保护作用”的假说提供实验支持。然而，在高山梨糖胁迫下，对照菌和酵母重组菌的生长情况无明显差异。

关键词: LEA3蛋白, PM2蛋白, 耐盐性, 酵母, 22-氨基酸基序

Abstract: Soybean PM2 protein belongs to the family of group 3 LEA (late embryogenesis abundant) proteins. In the present paper, yeast expression vectors encoding full-length PM2 protein and deleted polypeptides containing 22-mer motif were constructed. The yeast recombinants were obtained after transformation. Expression of PM2 and the deleted polypeptides of PM2A, PM2B and PM2C were identified in yeast lysis by SDS-PAGE and ESI-MS/MS or MALAI-TOF/TOF analysis. The growth performances of yeast recombinants and their control with empty vector were tested under normal, high salinity（1.5 mol·L^-1 NaCl）or high osmotic（2 mol·L^-1 sorbitol）conditions. The results showed that: 1) the expressions of PM2 protein and its 22-mer motif are not deleterious to the growth behavior of yeast recombinants under normal condition; 2) the recoveries of 4 recombinants are better than that of control with the empty vector under high salinity conditions, indicating that PM2 protein and its 22-mer motif could improve salt tolerance of yeast directly. The high salt tolerance of yeast conferred by 4 polypeptides are PM2C>PM2B≈PM2A≈PM2; 3) no growth difference was observed between the 4 recombinants and the control under high sorbitol stress. These results showed that 22-mer motif is a functional domain in PM2 protein and it is consist with the results described as in E. coli. All these results provide directly experimental support for the hypothesis that “LEA proteins may represent analogous protection to the high salinity stress in prokaryotes and eukaryotes” for the first time.

Key words: LEA3 proteins, PM2 protein, salt tolerance, yeast, 22-mer motif

中图分类号:

S565.1

刘昀;李冉辉;汪为茂;梁翔宇;郑易之*. 植物抗逆蛋白（LEA3）22-氨基酸耐盐结构域在酵母细胞中的鉴定[J]. 植物研究, 2009, 29(1): 74-79.

LIU Yun;LI Ran-Hui;WANG Wei-Mao;LIANG Xiang-Yu;ZHENG Yi-Zhi*. Functional Identification of Plant Stress-resistant Protein(LEA3) and Its 22-mer Motifs in Yeast[J]. Bulletin of Botanical Research, 2009, 29(1): 74-79.

[1]	赵玮, 齐燕妮, 张建平, 赵利, 王利民, 谢亚萍, 王斌, 张艳萍. 胡麻资源萌发期耐盐综合性评价[J]. 植物研究, 2019, 39(6): 955-963.
[2]	粟莉圆, 李孝哲, 陈淑雯, 张芳芳, 赵婷婷, 杨桂燕. 核桃JrGRAS2基因响应热胁迫的表达及功能分析[J]. 植物研究, 2018, 38(1): 125-131.
[3]	冯德明, 温佩颖, 赵畅, 杨远彪, 杨桂燕, 于丽丽, 高彩球. 刚毛柽柳ThDREB基因在酵母中的表达及抗逆能力分析[J]. 植物研究, 2017, 37(1): 63-68.
[4]	孟永斌1,2；徐蕾1,2；张子东1,2；路祺1,2；刘英1,2；聂思铭1,2；祖元刚1,2*. 纤维剩余物发酵制备乙醇工艺研究[J]. 植物研究, 2016, 36(1): 156-160.
[5]	臧丽娜；郑唐春；代丽娟；刘彩霞；曲冠证*. 白桦开花抑制因子BpFLC与BpSVP蛋白互作的结构域筛选与互作验证[J]. 植物研究, 2015, 35(6): 866-872.
[6]	纳晓莹1；王秀伟1*；徐浩玉1；乔丽娜1；王遂1；刘桂丰1；Kentbayev Yerzhan2；Kentbayeva Botagoz2. 4种桦树幼苗耐盐性分析与评价[J]. 植物研究, 2015, 35(6): 873-882.
[7]	王祥；张秀春；余乃通；梁洁；周朋；刘志昕. 香蕉束顶病毒nsp*酵母双杂交诱饵质粒的构建及自激活验证[J]. 植物研究, 2015, 35(5): 741-745.
[8]	马静1；曲春浦2；许志茹1,2*；杨传平2；刘关君2. 西伯利亚蓼铜伴侣蛋白与铜锌超氧化物歧化酶基因共转化烟草的耐盐性分析[J]. 植物研究, 2015, 35(2): 208-217.
[9]	谭老喜;张雨良;周朋;章绍延;王健华;张秀春;刘志昕*. 感染BBTV香蕉叶片cDNA文库的构建及评价[J]. 植物研究, 2013, 33(6): 713-717.
[10]	王艳君;王美燕;黄荣荣. 壳聚糖包衣对油菜种子萌发及幼苗耐盐性影响[J]. 植物研究, 2012, 32(6): 689-694.
[11]	张桦;张富春;张雨良;张博;陈全家. 新牧一号杂花苜蓿MvNHX*1基因的表达分析和提高烟草耐盐性的研究[J]. 植物研究, 2011, 31(5): 550-557.
[12]	林英超;杨蕾;赵晓菊;唐中华*. 盐胁迫条件下提高内源乙烯对拟南芥幼苗生长和根尖活性氧水平的影响[J]. 植物研究, 2010, 30(6): 703-707.
[13]	鲁振强;刘大丽;柳参奎. 过量表达水稻细胞质型OsAPXs基因提高转基因烟草的耐盐性[J]. 植物研究, 2010, 30(4): 448-454.
[14]	刘慧敏;朱月林*;陈磊. 组培条件下不同番茄品种及砧木自交系幼苗期硝酸盐耐性的比较[J]. 植物研究, 2007, 27(2): 175-181.
[15]	王振英, 彭永康, 郑坚瑜. 黑麦中与耐盐性相关的cDNA片段的克隆与序列分析[J]. 植物研究, 2005, 25(4): 427-431.

植物抗逆蛋白（LEA3）22-氨基酸耐盐结构域在酵母细胞中的鉴定

Functional Identification of Plant Stress-resistant Protein(LEA3) and Its 22-mer Motifs in Yeast

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价