北京林业大学学报2016，Vol.38Issue(7)：16-24,9.DOI:10.13332/j.1000-1522.20160027
白桦BpGT14基因启动子克隆及表达活性分析
Cloning and ;activity analysis of BpGT14 gene promoter in Betula platyphylla
摘要
Abstract
We cloned a 2 169 bp promoter sequence of BpGT14 gene from birch genomic DNA using the method of SiteFinding-PCR. The promoter sequence was analyzed by PLACE, and the result showed that this fragment contained promoter core elements and some elements which can respond to abiotic stress and hormones. Meanwhile, two important MYB transcription factor binding elements were found which regulate phenylpropanoid and lignin biosynthesis. To study the promoter activity, a 1 156 bp fragment was chosen to construct pBpGT14::GUS plant expression vector and transformed into tobacco. GUS staining proved that the promoter had high activity in stem segments. When the tobacco was treated with GA and H2 O2 at 4℃, the promoter had no significant response and the enzyme activity had a downward trend. In contrast, the promoter activity was significantly increased by ABA, NaCl, PEG and 37 ℃ treatment. Further transformation of birch cells using pBpGT14宜GFP plant expression vector indicated that the promoter had a similar response pattern to that of tobacco treated with abiotic stress and hormone except for a few time points. As the promoter was significantly and quickly responsive to drought stress, we have observed the GFP fluorescence protein in birch stem segments suspension cells transformed by GFP for 3 , 6, 12 and 24 h with PEG treatment. The results showed that BpGT14 promoter had activity in birch stem segments suspension cells and fluorescence can be observed in the suspension cells, especially in cell walls. Successful implementation of this study has important significance for analysis of gene regulation and function. Meanwhile, it provides a theoretical basis for gene promoter function studies of other woody plants.关键词
白桦糖基转移酶14基因/启动子克隆/报告基因/非生物胁迫Key words
glycosyltransferase14 gene/promoter clone/reporter gene/abiotic stress分类
农业科技引用本文复制引用
李蕾蕾,孙丰坤,李天宇,寇萍,詹亚光,曾凡锁..白桦BpGT14基因启动子克隆及表达活性分析[J].北京林业大学学报,2016,38(7):16-24,9.基金项目
中央高校基本科研业务费专项(2572014DA04)、国家自然科学基金项目(31200463、J1210053)。 （2572014DA04）
