QI Yingwei,LEI Qin,TIAN Jianwen,et al.Isolation and Characterization of the ‘Gala’ Apple MYB12 Gene Promoter[J].Northern Horticulture,2015,39(19):106-111.[doi:10.11937/bfyy.201519026]
‘嘎啦’苹果MYB12基因启动子的克隆与功能分析
- Title:
- Isolation and Characterization of the ‘Gala’ Apple MYB12 Gene Promoter
- 文献标志码:
- A
- 摘要:
- 以‘嘎啦’苹果基因组为模板，用PCR方法扩增得到苹果MYB12基因启动子（ProMYB12）。利用在线分析网站Plant CARE和PLACE对启动子上存在的顺式作用元件进行了预测。构建pCAMBI0390-ProMYB12-GUS植物瞬时表达载体并转化农杆菌，瞬时转染野生型番茄Micro-Tom （Lycopersion esculentum cv.Micro-Tom)用以研究启动子的启动活性。结果表明：克隆获得的启动子长度为1 290 bp。启动子序列中存在大量的顺式作用元件，既有转录必备的TATA-box和CAAT-box，也存在非生物胁迫和植物激素响应元件以及大量的光调控元件，此外还存在一些组织特异性表达元件。‘嘎啦’苹果MYB12启动子驱动的GUS基因在番茄的花和种子处有较高的表达量，表现出一定的组织特异性。
- Abstract:
- MYB12 promoter (ProMYB12) fragment was cloned and identified from genomic DNA of ‘Gala’ apple using the method of PCR.The sequence of this cloned promoter was analyzed by using the online database of Plant CARE and PLACE.The plant expression vector pCAMBI0390-ProMYB12-GUS was constructed and transformed into Agrobacteria GV3101.Through Arobacterium-mediated transformation，the constructed binary vector was introduced into wild type Micro-Tom （Lycopersion esculentum cv.Micro-Tom)tomato fruits，leaves and flowers in order to study the expression properties of this cloned promoter.The results showed that the cloned fragment was 1 290 bp long.Database searching showed that this fragment contained the essential elements such as TATA-box and CAAT-box，besides，there were several elements that responded to abiotic stress and hormones as well as many light responsive elements.Moreover，silico analysis of this isolated promoter sequence revealed the presence of some tissue specific cis-elements.Histochemical staining showed that GUS gene was highly expressed in the transgenic tomato flowers and around the tomato seeds which indicated the tissue specific characterization of the cloned promoter.
参考文献/References:
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备注/Memo
第一作者简介：戚英伟（1989-），男，硕士研究生，研究方向为果蔬贮藏与加工。E-mail：1533568558@qq.com.