葡萄逆境胁迫诱导启动子的克隆及表达分析  

Cloning and Expression Analysis of Grape's Stress Inducible Promoter

作　　者：吕兆勇[1] 赵春梅[2] 薛仁镐[2] 

机构地区：[1]聊城市人民医院,山东聊城252000 [2]青岛农业大学生命科学学院,山东青岛266109

出　　处：《华北农学报》2016年第1期77-82,共6页Acta Agriculturae Boreali-Sinica

基　　金：转基因生物新品种培育重大专项(2014ZX08010002-003-002);山东省自然科学基金项目(ZR2013CM025)

摘　　要：为了研究葡萄抗逆基因（CAN70200.1）的表达特性,采用PCR技术从葡萄中克隆了CAN70200.1基因上游一段长度为1 354 bp的启动子片段,命名为PCAN。采用Plant CARE和PLACE启动子在线预测工具分析表明,PCAN启动子序列具有CAAT-box、TATA-box基本的顺式作用元件和一些参与非生物胁迫、光和植物激素应答相关的顺式作用元件。为验证启动子的表达特性,将PCAN启动子连接到p CAMBIA1391Z载体GUS基因的上游,构建成植物表达载体p1391Z-CAN,并通过农杆菌介导法转化烟草,经PCR鉴定,获得转基因植株。对转基因烟草植株进行逆境胁迫处理发现,在干旱处理120 min后,PCAN启动子活性达到最强;而4℃低温处理30~60 min时,PCAN启动子活性达到最强,表明PCAN启动子具有低温和干旱胁迫诱导表达特性。To study the expression of grape stress tolerance gene（ CAN70200. 1）,a 1 354 bp promoter fragment（ named as PCAN） upstream of the gene CAN70200. 1 was isolated by using PCR technology. Promoter sequence was analyzed by the database of Plant CARE and PLACE. The result showed that the PCANsequence contained basic elements CAAT-box,TATA-box and some cis-acting elements that response to abiotic stresses,light and plant hormones. To verify the expression pattern of the promoter,the PCANfragment was fused with GUS reporter gene located on p CAMBIA1391 Z to construct a plant expression vector p1391Z-CAN,followed by transformation into tobacco by Agrobacterium-meditated method. The expression activity of PCANpromoter reached highest at 120 min after drought stress treatment or at 30- 60 min under 4 ℃ cold treatment condition,indicated that the PCANpromoter could express under the condition of treatments with cold and drought.

关 键 词：葡萄 PCAN启动子 胁迫诱导表达 GUS检测 

分 类 号：Q78[生物学—分子生物学] S565.1[农业科学—作物学]