| 沙冬青热胁迫相关蛋白基因AmHsa32超表达提高大肠杆菌的抗热性 |
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| 引用本文： | 王艳萍,刘美芹,师静,刘胜利,陈玉珍,卢有福.沙冬青热胁迫相关蛋白基因AmHsa32超表达提高大肠杆菌的抗热性[J].北京林业大学学报,2012,34(5):37-43. |
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| 作者姓名： | 王艳萍 刘美芹 师静 刘胜利 陈玉珍 卢有福 |
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| 作者单位： | 北京林业大学生物科学与技术学院,林木育种国家工程实验室;北京林业大学生物科学与技术学院,林木育种国家工程实验室;北京林业大学生物科学与技术学院,林木育种国家工程实验室;北京林业大学生物科学与技术学院,林木育种国家工程实验室;北京林业大学生物科学与技术学院,林木育种国家工程实验室;北京林业大学生物科学与技术学院,林木育种国家工程实验室 |
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| 基金项目： | 国家自然科学基金项目(30700559、30671476、31270737);北京市自然科学基金项目(6112016) |
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| 摘 要： | 热胁迫相关蛋白32（Heat stress associated protein 32，Hsa32）与植物的抗逆性密切相关。本文将从沙冬青低温干旱EST库中获得的热胁迫相关蛋白基因AmHsa32进行了非生物胁迫下的表达分析与转化大肠杆菌研究。生物信息学分析表明，AmHsa32的编码区全长858 bp，编码286个氨基酸，预测分子量约32 kD，与拟南芥HSA32亲缘关系较近。qRT PCR分析显示，AmHsa32在沙冬青幼苗中受热胁迫诱导后表达量迅速提高，1 h后达到对照的25倍；对其他非生物胁迫如低温、干旱及盐均有响应，表达量有不同程度的增加，说明AmHsa32对沙冬青非生物胁迫抗性的提高可能发挥着重要作用。将AmHsa32基因转化大肠杆菌，同野生菌相比，热胁迫（50 ℃）条件下，转基因大肠杆菌存活率明显提高。本研究结果表明，AmHsa32可用于通过转基因技术提高热敏感植物抗热性的研究。
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| 关 键 词： | 沙冬青 热胁迫 AmHsa32记基因 基因表达分析 转基因大肠杆菌 |
| 收稿时间： | 1900-01-01 |
Enhanced tolerance against heat stress of Escherichia coli cells by over-expressing an Ammopiptanthus mongolicus heat stress associated protein gene AmHsa32 |
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| WANG Yan-ping, LIU Mei-qin, SHI Jing, LIU Sheng-li, CHEN Yu-zhen, LU Cun-fu.Enhanced tolerance against heat stress of Escherichia coli cells by over-expressing an Ammopiptanthus mongolicus heat stress associated protein gene AmHsa32[J].Journal of Beijing Forestry University,2012,34(5):37-43. |
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| Authors: | WANG Yan-ping LIU Mei-qin SHI Jing LIU Sheng-li CHEN Yu-zhen LU Cun-fu |
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| Institution: | National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Biotechnology, Beijing Forestry University, 100083, P.R.China. |
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| Abstract: | The extremely summer high temperature in desert region makes the plant heat tolerant and a desirable model for revealing the mechanism of plant resistance to heat stress. In the present research, one gene designated as AmHsa32 encoding heat stress associated protein 32 was cloned by expressed sequence tags from cold- and drought-treated Ammopiptanthus mongolicus seedlings. Bioinformatic analysis showed that the ORF of AmHsa32 was composed of 858 base pairs,which encoded a heat stress associated protein with 286 amino acids and a molecular weight of 32 kD. AmHsa32 had a high affinity with heat stress associated protein 32 in Arabidopsis thaliana. qRT-PCR analysis showed that the expression level of AmHsa32 was 25-fold of control one hour later under heat stress and up-regulated under cold,drought,and salt stresses. Escherichia coli cells expressing AmHsa32 protein showed increased resistance to heat stress( 50 ℃ ) compared with control. We propose that AmHsa32 plays a protective role in cells under heat stress. This finding could potentially be used to improve the plant tolerance to heat stress via gene transfer. |
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| Keywords: | Ammopiptanthus mongolicus heat stress AmHsa32 gene gene expressing analysis transgenic Escherichia coli |
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