E3泛素连接酶基因CG4911敲除和功能的初步研究  

E3 Ubiquitin Ligase Gene CG4911 Gene Knock-out and Function Analysis

作　　者：叶晓蕾[1] 樊晓兰[1] 王爽爽[1] 张岩[2] 金龙金[3] 林鑫华[2] 

机构地区：[1]温州医学院眼视光学院,温州325027 [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,干细胞与发育生物学重点实验室,北京100101 [3]温州医学院检验医学院,生命科学学院,温州325035

出　　处：《中国细胞生物学学报》2012年第5期439-446,共8页Chinese Journal of Cell Biology

基　　金：国家重大科技计划专项课题(No.2011CB943900);温州医学院人才启动项目(No.QTJ08012);温州医学院科研基金重大项目(No.XNK07005);温州市科技局计划项目(No.Y20110154);温州医学院眼视光学院;医院院内课题(No.YNKT201113)资助项目~~

摘　　要：蛋白质的泛素化是一种重要的翻译后修饰过程,参与调控细胞周期、基因转录、信号转导、炎症反应和干细胞的维持等过程。泛素连接酶E3(ubiqutin ligase)是泛素化过程中关键酶。但许多E3基因在发育中的功能和作用机制还不明确。该研究以黑腹果蝇为模式动物,研究泛素连接酶家族一个重要基因CG4911的功能及分子机制。获得CG4911基因敲除果蝇,CG4911敲除果蝇纯合子可活。原位杂交结果显示,CG4911在胚胎发育早期表达。通过构建CG4911-pUAST-3HA重组子转染Hela细胞,确定CG4911定位于细胞质中,其表达并无修饰作用,并且过表达基因CG4911可导致背板发育缺陷。该研究首次获得了CG4911基因敲除果蝇和CG4911转基因果蝇,并初步探索了F-box基因CG4911的功能,为进一步阐明泛素连接酶的功能及分子机制提供了科学依据。Protein ubiquitination is an important post-translational modification process involved in cell cycle regulation,gene transcription,signal transduction,inflammation and stem cell maintenance.Ubiqutin ligase E3 is an essential enzyme and plays important role in ubiquitin-proteasome process.However,little is known about its function and molecular mechanism of E3 ligase during development.In this study,Drosophila melanogaster was used as a model to study the function and molecular mechanism of gene CG4911,which is one of the important ubiquitin ligases genes.We have generated mutant for CG4911 and construct CG4911-pUAST-V5 vector as well as its transgenic lines.The result showed that CG4911 mutant is viable.The protein of CG4911 localized in the cytoplasm and there was no modification happened with CG4911 when expressed in Hela cells.Furthermore,over expression of CG4911 leads to the notum developmental defects.In summary,this is the first time to get CG4911 knock out mutant and CG4911 transgenic fly.Also,it’s the first time to uncover a novel F-box gene CG4911 gene function in developmental contaxts.Our studies provide basis to further clarify the function of ubiquitin ligases and molecular mechanisms.

关 键 词：泛素连接酶基因 RNA干扰 原位杂交 果蝇 

分 类 号：R346[医药卫生—基础医学]