[1]叶展辉,郑易之,刘　昀.大豆PM2蛋白11氨基酸结构域的耐盐功能鉴定[J].深圳大学学报理工版,2006,23(4):362-367.
　YE Zhan-hui,ZHENG Yi-zhi,and LIU Yun.11-mer repeating region in soybean PM2 protein enhances salt tolerance of Escherichia coli[J].Journal of Shenzhen University Science and Engineering,2006,23(4):362-367.
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大豆PM2蛋白11氨基酸结构域的耐盐功能鉴定()
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《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第23卷
期数:
2006年4期
页码:
362-367
栏目:
土木建筑工程
出版日期:
2006-10-30

文章信息/Info

Title:
11-mer repeating region in soybean PM2 protein enhances salt tolerance of Escherichia coli
文章编号:
1000-2618（2006）04-0362-06
作者:
叶展辉;  郑易之;  刘　昀
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
Author(s):
YE Zhan-hui;  ZHENG Yi-zhi;  and LIU Yun
College o f Life Science Shenzhen University Shenzhen 518060 P. R. China
关键词:
LEA3蛋白; PM2蛋白; 11-氨基酸重复序列区; 结构域; 盐胁迫; 大肠杆菌; 大豆
Keywords:
LEA3 protein; PM2 protein; 11-mer repeating region;  structural region; salt tolerance; Escherichia coli; Soybean
分类号:
Q 943.2 786
文献标志码:
A
摘要:
大豆PM2蛋白属LEA3 (late embryogenesis abundant group3) 蛋白. 以含大豆PM2基因的重组载体为模板, PCR法扩增出可编码6拷贝11-氨基酸重复序列的基因片段PM2D. 构建pET28a /PM2D 重组载体, 并转化大肠杆菌得到重组菌BL21 /PM2D. SDS-PAGE 电泳结果表明, BL21 /PM2D 重组菌可被诱导表达相对分子质量约为15 000的蛋白条带, 与目的蛋白的理论相对分子质量(13 600) 接近. 利用DNASIS软件分析, PM2D 缺失短肽的亲水指数为1.12. 将重组菌BL21 /PM 2和BL21 /PM2D 分别涂布在含500mmo l /L N aC l和1 200mmol /L山梨糖的固体培养基上, 计算重组菌的存活率. 结果显示, 两种重组菌在含500mmo l /L N aC l培养基上的存活率分别为30.8%和26.0%, 均高于对照菌(BL21 /pET28a) 的存活率;在含1 200mmol /L山梨糖培养基上的存活率分别为59.7%和59.3%, 与对照BL21 /pET28a菌株的存活率相比无明显差异. PM2D短肽长度为PM2蛋白全长的1 /4, 但BL21 /PM2D重组菌的耐盐能力为BL21 /PM2重组菌的80%左右. 表明11-氨基酸重复序列区是PM2蛋白序列的一个耐盐结构域.
Abstract:
Soybean PM2 protein belongs to the family of group 3 LEA (late embryogenesis abundant) protein, which contains the conserved 11-mer repeating motif Nucleotide acid fragment PM2D encoding six copies of 11-aarepeating motif (aa292 ～ 404) was amplified by PCR reaction. The prokaryotic expression plasmid of pET28a /PM2D was constructed and then transformed into Escherichia coli strain to create recombinant BL21 /PM2D. The strains containing empty vector of pET28a and full-length PM2 gene were obtained as control, respectively. Total protein was isolated from the lysis of recombinant strain induced by IPTG. The result of SDS-PAGE analysis showed that a specific protein band of 15 000 appeared on the gel and it was close to the theoretic molecular weight (13 600) of PM2D polypeptide. This indicated that PM2D polypeptide could be expressed in recombinant BL21 /PM2D.The hydrophobicity index of PM2D polypeptide was calculated to be 1.12 by DNASIS software. On the medium supplemented with 500 mmol /L NaCl, the survival ratio of recombinant BL21 /PM 2 and BL21 /PM2D w as 30.8% and 26.0%, respectively, which w ere higher than 11.4% o f BL21 /pET28a. While on the medium containing 1 200mmol /L sorbitol, the survival ratio o f recombinant BL21 /PM2 and BL21 /PM2D w as 59.7% and 59.3%, respectively, showing no obvious difference compared to 54.9% o f BL21 /pET28a as control PM2D polypeptide is only a quarter of PM2 protein in length, but the survival ratio of BL21 /PM2D can reach to about 80% of BL21 /PM2. All these results indicate that 11-merrepea ting region is a functional region with salt-tolerance in PM2 (LEA3) protein.

相似文献/References:

[1]刘　昀,李冉辉,郑易之,等.大豆PM2蛋白及其结构域可提高烟草耐盐性[J].深圳大学学报理工版,2007,24(1):95.
　LIU Yun,LI Ran-hui,ZHENG Yi-zhi,et al.Soybean PM2 protein and its 22-mer region enhance salt tolerance of tobacco plants[J].Journal of Shenzhen University Science and Engineering,2007,24(4):95.

更新日期/Last Update: 2015-06-26