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当前位置: 首页 > 文章 > 日本结缕草ZjNAC1的克隆、转录激活活性、亚细胞定位及表达分析 中国草地学报 2019 (2) 1-8
Position: Home > Articles > Molecular Cloning, Transcriptional Activation, Subcellular Localization Analysis and Expression Characterization of ZjNAC1 from Zoysia japonica Chinese Journal of Grassland 2019 (2) 1-8

日本结缕草ZjNAC1的克隆、转录激活活性、亚细胞定位及表达分析

作  者：
滕珂;姜红岩;张蕊;檀鹏辉;岳跃森;范希峰;武菊英
单  位：
北京林业大学草坪研究所;北京草业与环境研究发展中心
关键词：
日本结缕草;NAC转录因子;转录激活;亚细胞定位;表达分析
摘  要：
采用RACE技术从日本结缕草中克隆得到ZjNAC1转录因子(GenBank No. MH428376),其开放阅读框为945bp,编码314个氨基酸。ZjNAC1编码的蛋白在N端有1个典型的NAM保守结构域,属于NAC转录因子家族。通过染色体步移的方法,获得ZjNAC1基因ATG上游1635bp序列,分析发现其包含响应脱落酸(ABA)、茉莉酸甲酯(MeJA)及逆境胁迫的作用元件。构建pGBKT7-ZjNAC1载体,转化Y2HGold酵母感受态细胞,发现ZjNAC1具有转录激活活性。农杆菌介导的35S-ZjNAC1-YFP载体注射本生烟草叶片瞬时表达结果显示,ZjNAC1定位于细胞核。实时荧光定量结果表明,ZjNAC1在日本结缕草根中表达量最高;此外,ZjNAC1的表达受10μmol/L MeJA和20%PEG4000处理的诱导,但受200μmol/L乙烯(ET)、10μmol/L ABA和300mmol/L NaCl处理的抑制。
译  名：
Molecular Cloning, Transcriptional Activation, Subcellular Localization Analysis and Expression Characterization of ZjNAC1 from Zoysia japonica
作  者：
TENG Ke;JIANG Hong-yan;ZHANG Rui;TAN Peng-hui;YUE Yue-sen;FAN Xi-feng;WU Ju-ying;Beijing Research and Development Center for Grass and Environment;Institute of Turfgrass Science,Beijing Forestry University;
关键词：
Zoysia japonica;;NAC transcription factor;;Transcriptional activity;;Subcellular localization;;Expression analysis
摘  要：
NAC transcription factors are a large class of unique transcription factors that play an important role in plant growth and development. In this study, the ZjNAC1(GenBank No. MH428376) was cloned from Zoysia japonica by RACE technology. The open reading frame of ZjNAC1 is 945 bp, encoding 314 amino acids. The ZjNAC1 protein has a typical NAM conserved domain at the N-terminus, belonging to the family of NAC transcription factors. The 1635 bp upstream sequence in front of the ATG of the ZjNAC1 transcription factor was obtained through Genome Walking. Bioinformatics analysis revealed that the sequence contained cis-elements responding to ABA、MeJA and stresses. The pGBKT7-ZjNAC1 vector was constructed and transformed into Y2 HGold yeast competent cells, and the result revealed that ZjNAC1 had transcriptional activation activity. The 35 S-ZjNAC1-YFP vector was transiently expressed in Nicotiana benthamiana leaves with agrobacterium-mediated method, and the results showed that ZjNAC1 was localized in the nucleus. Quantitative real-time PCR indicated that the expression of ZjNAC1 was abundantly accumulated in the root of Zoysia japonica. In addition, ZjNAC1 expression was induced by 10μmol/L MeJA and 20% PEG4000, but inhibited by 200μmol/L ET、10μmol/L ABA and 300 mmol/L NaCl. This study provides the basis for further exploration of the function of ZjNAC1.

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