用“增强子陷阱”技术构造并筛选果蝇UAS／GAL4系统中GAL4新品系及脑基因表达图谱数据库的开发

Genomic Screen for Expression Pattern in Adult Drosophila Brains In vivo by Enhancer Trap Method and Establishment of Expression Database

"增强子陷阱"技术是建立果蝇脑全基因组表达图谱及其数据库的重要方法.筛选获得新特异表达的GAL4品系,可为进一步研究果蝇脑神经在学习记忆功能提供强有力的基因工具.通过"增强子陷阱"技术来获得果蝇突变体,并与报告转基因果蝇(UAS-EGFP)杂交,用荧光显微镜观察成年果蝇脑内荧光分布,从而获得该突变体的脑基因表达图谱,在此基础上利用JavaScript来建立果蝇脑全基因组表达数据库.目前获得基因突变体果蝇2677种,大部分在果蝇脑中有表达,其中在果蝇嗅觉学习记忆相关脑区蘑菇体表达的基因有368个,且有部分基因特异地表达在某些传导通路上.这些果蝇基因突变体库及其表达图谱为进一步研究各基因的功能及作为遗传工具来研究各脑区结构和功能提供极大方便.

＂Enhancer trap＂ is an important technique to build and explore Drosophila brain gene expression pattern and its database. Large genome-wide screen for specific expression pattern in adult Drosophila brains by enhancer trap method provides fly tools for further understanding neural basis of learning and memory. Enhancer trap method was utilized to generate Drosophila P[Gal4] mutant library. Each mutant in library was crossed with GFP reporter flies （UAS-eGFP）; heads of the adult off-springs were dissected to obtain the whole brain which were examined under the epifluorescence microscope and expression pattern of each mutant were further analyzed. JavaScript was utilized to establish an expression pattern library. A total of 2 677 mutant lines of Drosophila were obtained by P-element induced mutagenesis and subjected to expression pattern analysis. Most of mutated genes express in adult fly. 368 mutated genes specific express in the mushroom body, which was reported to involve in Drosophila olfactory learning and memory, and some others specific express in other neuron circuitries. Drosophila mutant libraries and genome-wide expression pattern database was built by enhancer trap method, which as a tool will be utilized to understand the function of genes and particular neural circuitries in Drosophila in the future.