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番茄LeHsp110/ClpB基因的分子克隆及其对植物耐热性的影响

[Molecular cloning of tomato LeHsp110/ClpB gene and its effect on the thermotolerance in plant].

作者信息

Yang Jin-Ying, Sun Ying, Sun Ai-Qing, Yi Shu-Ying, Liu Jian

机构信息

College of Life Science of Shandong Normal University, Jinan 250014, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2006 Jan;22(1):52-7.

Abstract

The heat shock protein ClpB is a member of the Clp family and functions as molecular chaperones. ClpB is related to the acquired thermotolerance in organisms. A cDNA of 3144 bp was screened out of a tomato cDNA library. The polypeptide deduced from the longest ORF contains 980 amino acid residues, and was classified into HSP100/ClpB family based on the result of molecular phylogenesis analysis. Thus it was named as LeHSP110/ClpB according to its calculated molecular weight. LeHSP110/ClpB was characteristic of heat-inducibility but no constitutive expression, and was demonstrated to locate in chloroplastic stroma. An antisense cDNA fragment of LeHsp110/ClpB under the control of CaMV 35S promoter was introduced into tomato by Agrobacterium tumefactions-mediated method. At high temperature, the mRNA levels of LeHsp110/ClpB in antisense transgenic plants were lower than those in control plants. The PS II of transgenic plants is more sensitive to high temperature than that of control plants according to data of Fv/Fm. These results clearly showed that HSP110/ClpB plays an important role in thermotolerance of high plants.

摘要

热激蛋白ClpB是Clp家族的成员,作为分子伴侣发挥作用。ClpB与生物体后天获得的耐热性有关。从番茄cDNA文库中筛选出一个3144 bp的cDNA。从最长开放阅读框推导的多肽含有980个氨基酸残基,根据分子系统发育分析结果被归类到HSP100/ClpB家族。因此,根据其计算的分子量将其命名为LeHSP110/ClpB。LeHSP110/ClpB具有热诱导特性但无组成型表达,并且被证明定位于叶绿体基质中。通过根癌农杆菌介导的方法将CaMV 35S启动子控制下的LeHsp110/ClpB反义cDNA片段导入番茄。在高温下,反义转基因植株中LeHsp110/ClpB的mRNA水平低于对照植株。根据Fv/Fm数据,转基因植株的PS II对高温比对照植株更敏感。这些结果清楚地表明HSP110/ClpB在高等植物的耐热性中起重要作用。

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