High-Resolution CCD Camera and Luminometer for High-Throughput Luciferase-Based Analyses

该合同支持购买两个用于定位和测量荧光素酶活性的系统，该系统由融合到基因组启动子区域的荧光素酶基因组成的嵌合报告基因的表达而产生。这种融合现在通常被用作一种方便的、尽管是间接的基因调控措施，以及特定基因在生物体内表达的时机和定位。其中一个系统可以获取、处理和分析活性荧光素酶发出的光子。该系统将主要用于通过使用表达特定启动子-荧光素酶报告基因的转基因植物来跟踪体内基因表达的时间进程变化。第二个系统是一个高通量的96孔板读取器，旨在允许快速、自动化地测量细胞提取物中的发光。该仪器将用于使用各种细胞系的瞬时基因表达分析。这两项研究将由来自四个学术部门的五名调查人员使用。他们的研究包括：拟南芥生物钟的分子遗传学，植物糖感的遗传调控，玉米类黄酮生物合成和小分子运输的调控，植物细胞板形成的分子机制，以及哺乳动物细胞培养中生物钟基因转录的调控。此外，这些设备将用于研究生博士后的研究培训，并纳入现有的本科实验室技术课程。因此，该设备将加强所有级别的学生在现代活体成像和基于发光的技术方面的培训。

This award supports purchase of two systems to be used to localize and measure luciferase activity resulting from expression of chimeric reporter genes that consist of the luciferase gene fused to a promoter region of the genome. Such fusions are now routinely used as a convenient, though indirect, measure of gene regulation, and of the timing and localization of the expression of specific genes within an organism. One of the systems can acquire, process and analyze photons emitted by active luciferase. This system will be used primarily to follow time-course changes in gene expression in vivo, through the use of transgenic plants expressing specific promoter-luciferase reporter genes. The second system is a high throughput 96-well plate reader designed to allow rapid, automated measurements of luminescence in cell extracts. This instrument will be used for transient gene expression assays employing various cells lines. The two will be used by five investigators drawn from four academic departments. Their research includes studies of the molecular genetics of the circadian clock in Arabidopsis, the genetic regulation of sugar sensing in plants, the regulation of flavonoid biosynthesis and small molecular trafficking in maize, the molecular machinery of cell plate formation in plants, and the regulation of circadian clock gene transcription in mammalian cell culture. In addition, the equipment will be used in the research training of graduate postdoctoral students and incorporated into an existing undergraduate laboratory techniques course. The equipment will thus enhance student training at all levels in modern in vivo imaging and luminescence-based techniques.