果蝇Golgin-84在发育中的表达和功能研究  

Studies of Drosophila Golgin-84 Expression and Functions in Development

作　　者：侯国丽[1] 林鑫华[2] 吴一卉[2] 刁爱坡[1] 

机构地区：[1]天津科技大学生物工程学院,天津300457 [2]中国科学院动物研究所生物膜与膜生物工程国家重点实验室,北京100101

出　　处：《中国细胞生物学学报》2013年第5期622-629,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:31071181;31071284)资助的课题~~

摘　　要：高尔基体形态和结构的维持与真核细胞内的物质运输和细胞正常功能密切相关,高尔基体蛋白Golgin-84对维持高尔基体形态发挥了重要作用,但其在发育中的功能尚不清楚。为了研究Golgin-84在果蝇发育中的作用,成功制备并纯化了抗果蝇Golgin-84的多克隆抗体并成功构建了UAS-golgin-84-V5过表达和golgin-84 RNAi转基因果蝇,并对golgin-84在果蝇发育中的作用作了初步分析。蛋白免疫印迹和免疫荧光染色实验表明:制备的Golgin-84抗体能特异性识别重组和内源Golgin-84蛋白;Golgin-84与高尔基体蛋白GM130共定位;Golgin-84在果蝇不同发育阶段的各组织中广泛表达,在睾丸中表达较高。另外发现,选择性地在果蝇翅膀中抑制Golgin-84表达使得翅脉发育异常;过表达Golgin-84导致果蝇翅膀生长图式出现缺陷。该实验成功制备了果蝇Golgin-84抗体,获得了golgin-84转基因果蝇,并对golgin-84的功能作了初步的分析,为进一步阐明Golgin-84的作用机制奠定了基础。Abstract The structure of Golgi apparatus is closely related to membrane transport and normal cellular func- tion. It is already known that the cis-Golgi localized Golgin-84 plays an essential role in the maintenance of Golgi structure in mammalian cells. However, the function of Golgin-84 in development remains unclear. To examine the roles of Golgin-84 in development, we generated and purified the polyclonal antibody against Drosophila Golgin-84, and the UAS-golgin-84-V5 over-expression and golgin-84 RNAi transgenic Drosophila lines were generated. We pre- liminarily analyzed the function ofgolgin-84 in Drosophila melanogaster during wing development. Western blot and immunofluorescence staining showed that the purified antibody can recognize recombinant and endogenous Golgin-84 protein, and the antibody stained Golgin-84 co-localized with Golgi marker protein GM130 in S2 cells. Golgin-84 was ubiquitously expressed during development in Drosophila melanogaster and abundant in testis. Furthermore, inhibi- tion of Golgin-84 expression resulted in wing defects, and wing growth and patterning defects were observed in the Golgin-84 over-expressed Drosophila. In conclusion, we successfully generated the Golgin-84 antibody and transgenic lines in Drosophila, and examined the function of Golgin-84 in wing development. Our results laid the foundation to further study the physiological function and molecular mechanism ofgolgin-84.

关 键 词：Golgin-84 黑腹果蝇 抗体制备 过表达 

分 类 号：Q24[生物学—细胞生物学]