二色补血草LbGRP基因的克隆及抗逆能力分析

doi:10.3724/SP.J.1005.2010.00278

遗传 ›› 2010, Vol. 32 ›› Issue (3): 278-286.doi: 10.3724/SP.J.1005.2010.00278

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二色补血草LbGRP基因的克隆及抗逆能力分析

潘妍, 王玉成, 张大伟, 杨传平

东北林业大学林木遗传育种与生物技术教育部重点实验室, 哈尔滨 150040

收稿日期:2009-07-27 修回日期:2009-09-26 出版日期:2010-03-20 发布日期:2010-03-15
通讯作者: 杨传平 E-mail:yangchuanpingnefu@yahoo.com
基金资助:
黑龙江省国际合作项目(编号：WB07N02)和黑龙江省青年基金专项(编号：QC07C56)资助

Cloning and stress tolerance analysis of an LbGRP gene in Limonium bicolor

PAN Yan, WANG Yu-Cheng, ZHANG Da-Wei, YANG Chuan-Ping

Key Laboratory of Forest Tree Genetic Improvement and Biotechnology, Northeast Forestry University, Ministry of Education, Harbin 150040, China

Received:2009-07-27 Revised:2009-09-26 Online:2010-03-20 Published:2010-03-15
Contact: YANG Chuan-Beng E-mail:yangchuanpingnefu@yahoo.com

摘要/Abstract

摘要：

富含甘氨酸RNA结合蛋白(Glycine-rich RNA-binding proteins, GRP)是植物中重要的转录后调控蛋白, 在植物的生长发育及对胁迫的抗性调控等过程中起重要作用。文章从二色补血草(Limonium bicolor (Bunge) O.) cDNA文库中克隆出了富含甘氨酸RNA结合蛋白基因(LbGRP)的全长cDNA序列(GenBank登录号: GQ398238)。为了研究LbGRP的抗逆功能, 将其构建到酵母表达载体pYES2中, 转化到酿酒酵母INVSc1菌株中, 获得重组酵母INVSc1(pYES2-LbGRP), 同时将转空pYES2质粒的酵母INVSc1(pYES2)作为对照。对重组酵母INVSc1 (pYES2-LbGRP)和对照INVSc1(pYES2)进行NaCl、KCl、NaHCO₃、Na₂CO₃、干旱和冷冻胁迫, 比较它们在不同胁迫下的存活率。结果表明, INVSc1(pYES2-LbGRP)在各种胁迫下的存活率明显高于对照INVSc1(pYES2), 证明LbGRP基因具有抗NaCl、KCl、NaHCO₃、Na₂CO₃、干旱和冷冻等胁迫的能力, 推测该基因参与了二色补血草的抗逆调控过程。

关键词: 富含甘氨酸RNA结合蛋白, 抗逆, 酵母表达, 二色补血草

Abstract:

Glycine-rich RNA-binding proteins (GRP) are involved in post-transcriptional regulation, and play important roles in plant growth, development and stress response. In the present study, the full-length cDNA of a GRP gene (LbGRP, GenBank accession number: GQ398238) was cloned from a cDNA library of Limonium bicolor. To investigate the stress tolerance of LbGRP, the recombinant plasmid pYES2-LbGRP was constructed by inserting the LbGRP gene into the yeast expression vector pYES2. The recombinant plasmid pYES2-LbGRP was transformed into yeast Saccharomyces cerevisiae INVSc1, and the INVSc1 strain transformed with empty pYES2 was used as the control. The recombinant yeast INVSc1 harboring LbGRP (pYES2-LbGRP) and the control INVSc1 harboring empty pYES2 were treated with NaCl, KCl, NaHCO₃, Na₂CO₃, drought and frezzing, respectively, and their survial rates were compared under these stress conditions. The results showed that the survival rates of the recombinant yeast INVScl (pYES2-LbGRP) were higher than that of the control strain under these stress conditions, indicating that the LbGRP is tolerant to NaCl, KCl, NaHCO₃, Na₂CO₃, drought and frezzing stresses. These results suggested that LbGRP plays a role in stress tolerance of L. bicolor.

Key words: Limonium bicolor, glycine-rich RNA-binding protein, stress tolerance, yeast expression

潘妍，王玉成，张大伟，杨传平. 二色补血草LbGRP基因的克隆及抗逆能力分析[J]. 遗传, 2010, 32(3): 278-286.

BO Yan, WANG Yu-Cheng, ZHANG Da-Wei, YANG Chuan-Beng. Cloning and stress tolerance analysis of an LbGRP gene in Limonium bicolor[J]. HEREDITAS, 2010, 32(3): 278-286.

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二色补血草LbGRP基因的克隆及抗逆能力分析

Cloning and stress tolerance analysis of an LbGRP gene in Limonium bicolor

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