Cloning of a cytosolic ascorbate peroxidase gene from Lycium chinense Mill. and enhanced salt tolerance by overexpressing in tobacco.
Cloning of a cytosolic ascorbate peroxidase gene from Lycium chinense Mill. and enhanced salt tolerance by overexpressing in tobacco.
复制标题
从枸杞中克隆胞质抗坏血酸过氧化物酶基因。
DOI：
10.1016/j.gene.2014.03.061
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发表时间：
2014
期刊：
影响因子：
3.5
通讯作者：
Yurong Wang
中科院分区：
文献类型：
--
作者：
Guangxi Wu;Gang Wang;J. Ji;Hailing Gao;Wenzhu Guan;Jiang Wu;Chunfeng Guan;Yurong Wang
为了研究胞浆抗坏血酸过氧化物酶（APX）在活性氧（ROS）清除系统中的生理意义，从枸杞（Lycium chinense Mill.）利用同源克隆的方法，研究了LmAPX在盐胁迫下的表达。经测序及相关分析，LmAPX cDNA序列长965 bp，开放阅读框（ORF）为750 bp，编码250个氨基酸。将LmAPX序列亚克隆到原核表达载体pET 28a中，重组蛋白在大肠杆菌中得到高效表达。Southern杂交分析结果表明，该基因的三个插入存在于烟草基因组中编码LmAPX。与对照（野生型和空载体对照）相比，转LmAPX基因植株在相同盐胁迫下表现出较低的过氧化氢（H2O2）含量和较高的抗坏血酸过氧化物酶活性、脯氨酸含量和净光合速率（Pn）。这些结果表明，LmAPX基因的过表达可以减少由盐胁迫引起的ROS的产生，保护植物免受氧化胁迫。
To evaluate the physiological importance of cytosolic ascorbate peroxidase (APX) in the reactive oxygen species (ROS)-scavenging system, a full-length cDNA clone, named LmAPX, encoding a cytosolic ascorbate peroxidase was isolated from Lycium chinense Mill. using homologous cloning, then the expression of LmAPX under salt stress was investigated. After sequencing and related analysis, the LmAPX cDNA sequence was 965 bp in length and had an open reading frame (ORF) of 750 bp coding for 250 amino acids. Furthermore, the LmAPX sequence was sub-cloned into prokaryotic expression vector pET28a and the recombinant proteins had a high expression level inEscherichia coli. Results from a southern blot analysis indicated that three inserts of this gene existed in the tobacco genome encoding LmAPX. Compared with the control plants (wild-type and empty vector control), the transgenic plants expressing the LmAPX gene exhibited lower amount of hydrogen peroxide (H2O2) and relatively higher values of ascorbate peroxidase activity, proline content, and net photosynthetic rate (Pn) under the same salt stress. These results suggested that overexpression of the LmAPX gene could decrease ROS production caused by salt stress and protect plants from oxidative stress.
影响因子：
7.4
作者：
Asada, Kozi
通讯作者：
Asada, Kozi