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Position: Home > Articles > Transcriptional Activation Identification of the Transcription Factor HcSCL13 from Halostachys caspica and the Construction of Plant Expression Vector Biotechnology Bulletin 2014,0 (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

作  者:
周莲洁;张富春;王艳
单  位:
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
关键词:
盐穗木;转录因子;HcSCL13;转录激活;植物表达载体
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。
译  名:
Transcriptional Activation Identification of the Transcription Factor HcSCL13 from Halostachys caspica and the Construction of Plant Expression Vector
作  者:
Zhou Lianjie;Zhang Fuchun;Wang Yan;Xinjiang Key Laboratory of Biological Resources and Genetic Engineering,College of Life Science and Technology,Xinjiang University;
关键词:
Halostachys caspica Transcription factor HcSCL13 Transcriptional activation Plant expression vector
摘  要:
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.

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