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1885 bp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T1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2 h,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 iso-late and characterize the GmERF9 promoter ( GmERF9P) from soybean. The GmERF9P was amplified with PCR from the genome DNA of soybean,which had a length of 1885 bp. Sequence analysis indicated that GmERF9P contained a number of stress-related elements. The GmERF9P was cloned into plant expression vector of pCAM-BIA1301 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 in-creased under cold treatment for 2 h,which proved that GmERF9P was an efficient cold-inducible promoter.
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