欢迎访问《中国农学通报》，
中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (27): 178-182.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

维管特异表达启动子BSP的克隆与活性检测

刘玲玲 张金文   

  • 收稿日期:2011-11-28 修回日期:2012-02-27 出版日期:2012-09-25 发布日期:2012-09-25

Vascular-specific Expression and Cloning of BSP Promoter Activity Assay

  • Received:2011-11-28 Revised:2012-02-27 Online:2012-09-25 Published:2012-09-25

PDF (PC)

707

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-BA 0.5 mg/L+NAA 0.05 mg/L+Kan 100 mg/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OLYPUS BX51/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。

关键词: 遗传多样性, 遗传多样性

Abstract:

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.