盐穗木转录因子HcSCL13转录激活的体外鉴定及植物表达载体的构建

生物技术通报 ›› 2014, Vol. 0 ›› Issue (5): 76-82.

盐穗木转录因子HcSCL13转录激活的体外鉴定及植物表达载体的构建

周莲洁张富春王艳

(新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室，乌鲁木齐 830046)

收稿日期:2013-11-17 出版日期:2014-05-23 发布日期:2014-05-24
作者简介:周莲洁，女，硕士研究生，研究方向：植物抗逆的生理生化与分子机制；E-mail：zhoulianjie90@163.com
基金资助:
"973"计划前期研究专项(2012CB722204)，新疆维吾尔自治区高校科研计划科学研究重点项目(XJEDU2012102)，新疆大学博士启动基金项目(XJEDU2009S04)，新疆生物资源基因工程重点实验室开放课题(XJDX0201-2013-02)

Transcriptional Activation Identification of the Transcription Factor HcSCL13 from Halostachys caspica and the Construction of Plant Expression Vector

Zhou Lianjie Zhang Fuchun Wang Yan

(Xinjiang Key Laboratory of Biological Resources and Genetic Engineering，College of Life Science and Technology，Xinjiang University，Urumqi 830046)

Received:2013-11-17 Published:2014-05-23 Online:2014-05-24

摘要/Abstract

摘要： 通过同源重组的方法将盐穗木盐响应GRAS家族转录因子HcSCL13基因(GenBank登录号：KC68640)构建至酵母双杂交诱饵表达载体pGBKT7上，转化酵母菌Y2HGold，通过自激活性和毒性检测确定该转录因子的体外激活能力。试验表明，重组菌株pGBKT7-HcSCL13/Y2HGold对宿主菌无毒性，且该菌株表达的融合蛋白能够激活报告基因的表达，说明HcSCL13具有转录激活域，是一类转录因子。成功构建了HcSCL13基因的植物表达载体和亚细胞定位表达载体，为进一步在体内研究该基因的耐盐功能奠定了基础。

关键词: 盐穗木, 转录因子, HcSCL13, 转录激活, 植物表达载体

Abstract: HcSCL13 gene of the GRAS family from Halostachys caspica(GenBank accession number：KC68640)was amplified and ligated to the pGBKT7 vector by homologous recombination to construct the bait vector pGBKT7-HcSCL13, and then transformed into the competent yeast Y2HGold. Self-activation of recombinant bait vector in yeast two-hybrid system was measured. The results showed that the bait protein had not toxic to Y2HGold and could activate all the reporter genes. Therefore, we proposed HcSCL13 was a transcription factor with a transcriptional activation domain. The plant over-expression vector and subcellular localization expression vector were successfully constructed for further studying its biological function.

Key words: Halostachys, Caspica Transcription factor HcSCL13, Transcriptional activation, Plant expression vector

周莲洁, 张富春, 王艳. 盐穗木转录因子HcSCL13转录激活的体外鉴定及植物表达载体的构建[J]. 生物技术通报, 2014, 0(5): 76-82.

Zhou Lianjie, Zhang Fuchun, Wang Yan. Transcriptional Activation Identification of the Transcription Factor HcSCL13 from Halostachys caspica and the Construction of Plant Expression Vector[J]. Biotechnology Bulletin, 2014, 0(5): 76-82.

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