维管特异表达启动子BSP的克隆与活性检测  

Vascular-specific Expression and Cloning of BSP Promoter Activity Assay

作　　者：刘玲玲[1] 张金文[2] 

机构地区：[1]定西师范高等专科学校,甘肃定西743000 [2]甘肃农业大学,兰州730070

出　　处：《中国农学通报》2012年第27期178-182,共5页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金(30270843);甘肃省自然科学基金(ZS991-A21-029-N);定西市科技攻关计划项目[2005-1-15;定市科发(2005)51号文件]

摘　　要：为了研究维管特异表达启动子BSP的活性和组织特异性,从杨树基因组中克隆得到维管特异表达启动子BSP,利用该启动子与绿色荧光蛋白(GFP)报告基因构建植物表达载体,采用基因枪法转化烟草叶片,并在高渗培养基上培养过夜,后转移到诱导培养基上(MS+6-BA0.5mg/L+NAA0.05mg/L+Kan100mg/L)培养生根并长到一定高度后,剪下已形成维管组织的叶脉和根,制成切片,然后在OLYPUSBX51/BX52型荧光显微镜下用蓝光激发,观察各个组织细胞中的绿色荧光,同时以未转化的烟草材料做为对照。结果表明:BSP启动子所驱动的GFP基因在烟草的维管组织细胞中得到了高水平的表达,而在烟草的其他组织及对照的任何组织中几乎不表达。通过GFP在烟草维管组织中的瞬时表达,表明了该启动子具有维管组织特异性表达活性特点。The vascular-specific expression promoter BSP was cloned from the poplar genome in order to study the activity and tissue specificity, and the plant expression vector was constructed with the promoter and the green fluorescent protein （GFP） reporter gene, then transformed the tobacco leaves using the biolistic method. The trans-formed tobacco were cultured in hypertonic medium overnight, then transferred to the inducted media （MS＋6-BA 0.5 mg/L＋NAA 0.05 mg/L ＋Kan 100 mg/L） to take root and when they grown to a certain height, the section were prepared using the vein and root which had formed the vascular tissue. The green fluorescence of each tissue was observed by the fluorescence microscope of OLYPUS BX51/BX52 with the blue excitation, at the same time the untransformed tobacco was as control. Results indicated that the GFP gene driving by BSP promoter had been a high level of expression in vascular tissue cells in comparing with the other tissue and the control group. The transient expression of GFP in tobacco vascular tissues suggested that the promoter had the activity of vascular tissue-specific expression.

关 键 词：维管特异表达启动子 报告基因 表达载体 GFP瞬时表达 

分 类 号：Q78[生物学—分子生物学]