大豆低温诱导启动子GmERF9P的克隆及活性鉴定  

Cloning and Activity Analysis of Cold Induced GmERF9P Promoter from Soybean

作　　者：翟莹[1] 张军[2] 任巍巍[1] 张闯[1] 赵艳[1] 张梅娟[1] ZHAI Ying ZHANG Jun REN Weiwei ZHANG Chuang ZHAO Yan ZHANG Meijuan(College of Life Science and Agro-Forestry,Qiqihar University,Qiqihar 161006,China Heilongjiang Institute of Veterinary Science,Qiqihar 161005,China)

机构地区：[1]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006 [2]黑龙江省兽医科学研究所,黑龙江齐齐哈尔161005

出　　处：《华北农学报》2017年第5期13-18,共6页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31301335);黑龙江省教育厅科学技术研究项目(12541889);黑龙江省自然科学基金项目(C201458);黑龙江省普通本科高等学校青年创新人才培养计划(UNPYSCT_2016090)

摘　　要：为获得低温诱导基因GmERF9启动子,并分析该启动子的功能,利用PCR技术从大豆叶片基因组DNA中克隆1885bp的GmERF9启动子序列GmERF9P。序列分析表明,GmERF9P序列中含有多种与逆境相关的顺式作用元件。将GmERF9P构建到植物表达载体pCAMBIA1301上并转化烟草。通过PCR检测共获得6株T_1阳性转基因烟草株系。对野生型烟草和转基因烟草进行低温处理2h,通过GUS组织化学染色和实时荧光定量PCR检测GUS基因的表达量。结果显示GmERF9P在低温处理下能够提高GUS基因的表达量,具有低温诱导启动活性。Promoters play a key role in the regulation of gene expression. The objective of this study is to isolate and characterize the Gm ERF9 promoter（ GmERF9P） from soybean. The GmERF9P was amplified with PCR from the genome DNA of soybean,which had a length of 1 885 bp. Sequence analysis indicated that GmERF9P contained a number of stress-related elements. The GmERF9P was cloned into plant expression vector of p CAMBIA1301 and transformed into tobacco NC89. The regenerated plants were analyzed by PCR and showed that the promoter fragments were integrated into six genomes of tobacco plants. Histochemical staining of GUS activity and Real-time fluorescent quantitative PCR of the transgenic plants showed that the expression of GUS gene was increased under cold treatment for 2 h,which proved that GmERF9P was an efficient cold-inducible promoter.

关 键 词：大豆 GmERF9P 启动子 低温诱导 转基因烟草 

分 类 号：Q78[生物学—分子生物学] S565.03[农业科学—作物学]