《植物生理学报》 2015, 51(7): 1157-1166

小桐子转录因子CBF基因的克隆及酵母表达载体的构建

王海波^1,2, 王莎莎¹, 邹竹荣¹, 龚明^1,*

¹云南师范大学生命科学学院, 生物能源持续开发利用教育部工程研究中心, 云南省生物质能与环境生物技术重点实验室, 昆明650500; ²曲靖师范学院生物资源与环境科学学院, 云南曲靖655011

通信作者：龚明；E-mail: gongming63@163.com；Tel: 0871-65516516

摘　要：

高等植物不依赖ABA的低温信号转导途径中, 存在于细胞膜上的低温感受器通过ICE-CBF-COR级联响应将外界低温信号传导至胞内, CBF转录因子是承接上游ICE类转录激活因子与下游COR类低温响应基因的关键环节。本文利用前期获得的小桐子低温锻炼转录组数据, 克隆到了一条小桐子CBF基因的cDNA序列, 命名为JcCBF1 (GenBank登录号KJ670147.1)。结果表明, 该cDNA序列全长1 034 bp, 完整开放阅读框729 bp, 编码242个氨基酸, 分子量为26.6 kDa, 理论等电点为9.38, 具有CBF家族典型的AP2基序, 同时构建了其酵母表达载体, 为小桐子CBF转基因功能验证奠定基础。

关键词：小桐子; 转录因子; CBF; 不依赖ABA的低温信号转导; 基因克隆; 表达载体构建

收稿：2015-04-14   修定：2015-06-08

资助：国家自然科学基金(31260064、31460059、31460179和31460182)和云南省教育厅科研基金重大专项项目(ZD2010004)。

Molecular Cloning and Yeast Expression Vector Construction of the Transcription Factor Encoding CBF from Jatropha curcas

WANG Hai-Bo^1,2, WANG Sha-Sha¹, ZOU Zhu-Rong¹, Gong Ming^1,*

¹School of Life Sciences, Yunnan Normal University, Engineering Research Center of Sustainable Development and Utilization of Biomass Energy of Ministry of Education, Key Laboratory of Biomass Energy and Environmental Biotechnology of Yunnan Province, Kunming 650500, China; ²College of Biological Resource and Environmental Science, Qujing Normal University, Qujing, Yunnan 655011, China

Corresponding author: Gong Ming; E-mail: gongming63@163.com; Tel: 0871-65516516

Abstract:

Low temperature receptor present on the cell membrane can transfer environmental low temperature signal to inner of cells through cascade response of ICE-CBF-COR in ABA-independent cold signal transduction pathway. CBF transcription factor is a key step to connect the upstream of ICE activating transcription factor and downstream of COR low temperature responsive genes. A gene, named JcCBF1 (GenBank accession KJ670147.1) was cloned based on the chilling hardening transcriptome of Jatropha curcas. The full-length cDNA of JcCBF1 was 1 034 bp, containing a 729 bp open reading frame. The ORF encoded a 242 amino acid polypeptide with the molecular weight of 26.6 kDa and the pI value of 9.38. The JcCBF1 protein encompasses conserved motif of AP2. Expression vectors of Saccharomyces cerevisiae were constructed, in order to lay the foundation for CBF transgenic verification in J. curcas.

Key words: Jatropha curcas; transcription factor; CBF; ABA-independent cold signal transduction pathway; gene cloning; expression vector construction