[1]赵艳玲,姚婕.巨尾桉铜分子伴侣EuCCS基因的克隆及表达分析[J].华侨大学学报(自然科学版),2017,38(3):374-378.[doi:10.11830/ISSN.1000-5013.201703016]
 ZHAO Yanling,YAO Jie.Cloning and Expression of Copper Chaperone for Superoxide Dismutase Gene in Eucalyptus grandis×E. ophylla[J].Journal of Huaqiao University(Natural Science),2017,38(3):374-378.[doi:10.11830/ISSN.1000-5013.201703016]
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巨尾桉铜分子伴侣EuCCS基因的克隆及表达分析()
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《华侨大学学报(自然科学版)》[ISSN:1000-5013/CN:35-1079/N]

卷:
第38卷
期数:
2017年第3期
页码:
374-378
栏目:
出版日期:
2017-05-20

文章信息/Info

Title:
Cloning and Expression of Copper Chaperone for Superoxide Dismutase Gene in Eucalyptus grandis×E. ophylla
文章编号:
1000-5013(2017)03-0374-05
作者:
赵艳玲 姚婕
华侨大学 化工学院, 福建 厦门 361021
Author(s):
ZHAO Yanling YAO Jie
College of Chemical Engineering, Huaqiao University, Xiamen 361021, China
关键词:
巨尾桉 铜分子伴侣 克隆 原核表达 聚合酶链式反应
Keywords:
Eucalyptus grandis×E. ophylla copper chaperone for superoxide dismutase clone prokaryotic expression polymerase chain reaction
分类号:
Q943.2
DOI:
10.11830/ISSN.1000-5013.201703016
文献标志码:
A
摘要:
以巨尾桉为研究对象,克隆得到巨尾桉铜分子伴侣(CCS)基因(GenBank注册号为KJ755351.1),生物信息学分析表明:EuCCS蛋白定位于叶绿体,与龙眼同源基因的一致性达到99%.巨尾桉CCS的原核表达载体pET-EuCCS在大肠杆菌细胞内可以稳定表达,超氧化物歧化酶(SOD)活检测结果表明:转化菌株的SOD酶活性比对照菌株高,说明外源基因EuCCS具有生物学活性.利用荧光定量聚合酶链式反应(PCR)技术分析CCS在巨尾桉中的表达特性,结果表明:在植株生长旺盛的部位,CCS表达量较大,如植株幼叶,随植株叶片慢慢衰老,CCS的表达量也随之下降;在组培苗中,叶片的CCS表达量最大,其次是茎.
Abstract:
The major pathway for activation of Cu/ZnSOD requires the CCS copper chaperone to insert copper and activate Cu/ZnSOD through oxidation of an intramolecular disulfide. The EuCCS gene is cloned by RT-PCR from Eucalyptus grandis×E. ophylla(GenBank Accession Number: KJ755351.1).Boinformatics analysis suggests that CCS is located in the chloroplast and the evolutionary distance between eucalyptus and longan is quite close with the identity of 99%.The strain prokaryotic expression vector pET-EuCCS can be expressed stably in E. coli, and the superoxide dismutase enzyme activityis higher than control indicating that the cloned EuCCS gene could activate Cu/ZnSOD.Besides, the expression characteristics of eucalyptus CCS gene are analyzed by quantitative PCR.And the results show that the EuCCS quantity of the young leaves is relatively higher than that of the old one to greenhouse seedling and for tissue culture seedling the EuCCS is expressed higher in leaves than stem and root.

参考文献/References:

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相似文献/References:

[1]赵艳玲,周利建.巨尾桉胞质EuCuZnSOD基因的克隆与原核表达[J].华侨大学学报(自然科学版),2015,36(6):693.[doi:10.11830/ISSN.1000-5013.2015.06.0693]
 ZHAO Yanling,Zhou Lijian.Cloning and Expression of the Cytosolic Copper/Zinc Superoxide Dismutase Gene in Eucalyptus grandis×E. ophylla[J].Journal of Huaqiao University(Natural Science),2015,36(3):693.[doi:10.11830/ISSN.1000-5013.2015.06.0693]

备注/Memo

备注/Memo:
收稿日期: 2016-05-18
通信作者: 赵艳玲(1978-),女,助理研究员,博士,主要从事植物分子生物学的研究.E-mail:zhaoyl@hqu.edu.cn.
基金项目: 国家自然科学基金资助项目(31300497)
更新日期/Last Update: 2017-05-20