果蝇翅芽特异表达Gal4的筛选及活力检测

Screening and Activity Assays of the Drosophila Wing-specific Gal4

筛选并鉴定在果蝇翅芽不同区域特异表达的Gal4,可为以果蝇翅为模型的相关研究提供更多的遗传工具。本研究对Bloomington Drosophila Stock Center(BDSC)未见详细报道的Gal4品系进行筛选,将其中12个候选Gal4品系的雄虫与UAS-GFP基因型的处女蝇分别杂交,检测Gal4在子代幼虫翅芽的表达位置。经筛选重点关注3个Gal4,并以翅芽特异隔间标记物为参照,精确表征Gal4在翅芽的表达部位。结果显示:c563a-Gal4在前隔间翅囊区边缘及部分铰链区表达;c804-Gal4在围绕翅囊的环形区域以及A/P边界的细胞条带处表达;GMR11F02-Gal4在整个翅囊区表达;利用G-TRACE技术对上述Gal4的细胞发育谱系进行追踪,进一步揭示了翅芽发育历程中开启Gal4表达细胞的子代细胞分布区域;同时,通过驱动特定靶基因myc的表达,检测Gal4激活靶基因表达的活力。结果表明,3种Gal4都可以有效激活靶基因myc。其中,GMR11F02-Gal4的活力最强,c563a-Gal4最弱。本研究为翅芽相关研究提供新的遗传工具选项。

Screening Gal4 with specific expression patterns in Drosophila wing disc can provide more genetic tools for using Drosophila wing as a research model.In this study,12 Gal4 stocks from the Bloomington Drosophila Stock Center(BDSC)that have not been reported in detail in previous studies were screened.Males in the stock were crossed with virgin flies of the UAS-GFP stock,and the expression pattern of Gal4 in the wing disc of progenies was detected and identified.After screening,we focused on three Gal4 and accurately characterized their expression patterns according to wing disc-specific markers.We found that c563a-Gal4 is expressed at the edge of the anterior wing pouch region and part of the hinge region in the anterior compartment;c804-Gal4 is expressed in the ring surrounding the wing pouch and a cell stripe adjacent to the A/P boundary;GMR11F02-Gal4 is expressed in the entire wing pouch.We further used the G-TRACE technology to trace the cell lineage of Gal4 to determine the distribution domain of progeny cells that turned on Gal4 expression during wing disc development.Furthermore we expressed myc under the control of these three Gal4 drivers to examine the Gal4 activity.The results showed that these three Gal4 are sufficient to activate the target gene myc.The GMR11F02-Gal4 activity is the strongest,while the c563a-Gal4 activity is the lowest.Hence,this study provides new genetic tools for wing disc research.