大豆转录因子GmERF5 启动子的克隆及活性分析

生物技术通报 ›› 2014, Vol. 0 ›› Issue (1): 100-104.

大豆转录因子GmERF5 启动子的克隆及活性分析

翟莹¹,张军²,赵艳¹,孙天国¹,姚雅男³,杨晓杰¹

1. 齐齐哈尔大学生命科学与农林学院遗传重点实验室，齐齐哈尔 161006
2. 黑龙江省兽医科学研究所，齐齐哈尔 161006
3. 长春农业博览园，长春 130117

收稿日期:2013-09-06 出版日期:2014-01-23 发布日期:2014-01-23
作者简介:翟莹，女，博士，讲师，研究方向: 大豆分子育种；E-mail: fairy39809079@126.com
基金资助:
齐齐哈尔大学青年教师科研启动支持计划项目(2012k-M20)

Cloning and Activity Analysis of Soybean GmERF5 Promoter

Zhai Ying¹, Zhang Jun², Zhao Yan¹, Sun Tianguo¹, Yao Yanan³, Yang Xiaojie¹

1. College of Life Science and Agro-forestry，Qiqihaer Univesity，Key Laboratory of Genetics，Qiqihaer 161006;
2. Heilongjiang Institute of Veterinary Science，Qiqihaer 161006;
3. Changchun Agriculture Expo Garden，Changchun 130117

Received:2013-09-06 Published:2014-01-23 Online:2014-01-23

摘要/Abstract

摘要： 乙烯响应因子(Ethylene-responsive factors，ERFs) 广泛参与植物对生物和非生物胁迫的应答。从大豆吉林32 的#br#基因组DNA 中分离到GmERF5 的启动子片段，全长1 924 bp 。该区段含有9 种与逆境相关的顺式作用元件，即G-box 、GT-1 、#br#LTRE-1 、W-box 、TL1 、DPBF 、MYB 、MYC 和BIHD10S 。将该启动子构建到植物表达载体pCAMBIA1301 上与葡萄糖苷酸酶(GUS)#br#基因融合，注射法转化烟草叶片，并进行干旱、高盐和低温处理，通过GUS 组织化学染色和GUS 荧光值的测定分析启动子的启动#br#活性。结果表明，在未处理条件下，该启动子能驱动下游GUS 基因的表达。干旱和低温处理10 h 能明显增强GmERF5P 的启动活性。

关键词: 大豆, GmERF5, GUS 活性, 启动子, 逆境诱导

Abstract: ERF transcription factor family is extensively involved in the biotic and abiotic responses. The GmERF5 promoter(1 924 bp) was isolated from soybean genome using PCR. Sequence analysis indicated that GmERF5P contains several stress-induced elements: G-box,GT-1, LTRE-1, W-box, TL1, DPBF, MYB, MYC and BIHD10S. GmERF5P was subcloned into pCAMBIA1301 vector to drive the GUS gene to express in tobacco leaves by Agrobacterium-mediated injection transformation. The activity of GmERF5P was analyzed using histochemistry staining and GUS fluorescence intensity analysis. The results indicated that GmERF5P activated expression of GUS report gene under normal growing conditions. The activity of GmERF5P was up-regulated after the treatments of drought and low temperature for 10 h.

Key words: Soybean, GmERF5P, GUS activity, Promoter, Stress-induced

翟莹,张军,赵艳,孙天国,姚雅男,杨晓杰. 大豆转录因子GmERF5 启动子的克隆及活性分析[J]. 生物技术通报, 2014, 0(1): 100-104.

Zhai Ying, Zhang Jun, Zhao Yan, Sun Tianguo, Yao Yanan, Yang Xiaojie. Cloning and Activity Analysis of Soybean GmERF5 Promoter[J]. Biotechnology Bulletin, 2014, 0(1): 100-104.

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