人源胞红蛋白转基因秀丽线虫的制备与鉴定  

Construction and characterization of transgenic C. elegans expressing human cytoglobin

作　　者：赵娜[1,2] 任长虹[2] 张继业[1] 庞剑会[1] 刘虎岐[1] 张成岗[1,2] 

机构地区：[1]西北农林科技大学生命科学学院,陕西杨凌712100 [2]军事医学科学院放射与辐射医学研究所,蛋白质组学国家重点实验室,北京100850

出　　处：《军事医学科学院院刊》2009年第4期304-307,321,共5页Bulletin of the Academy of Military Medical Sciences

基　　金：国家973计划项目(2006CB504100);国家科技重大专项课题(2009ZX09103-616);国家自然科学基金项目(30771230,30600365);中国博士后科学基金特别资助基金(200801495);中国博士后科学基金(20060400051)资助

摘　　要：目的:为了以秀丽线虫为模式生物研究人源胞红蛋白(human cytoglobin,hCGB)的生理功能,构建hCGB的秀丽线虫表达载体,并通过显微注射的方法制备转基因线虫。方法:将通过RT-PCR获得的hCGB的cDNA序列亚克隆到pFX-unc-119表达载体中,通过显微注射的方法获取染色体外转基因线虫,采用紫外线(UV)照射的方法,筛选获得整合到染色体中的株系,并采取Western印迹和免疫组织化学法鉴定外源基因的表达。结果与结论:成功地构建了hCGB基因的表达载体并获得了3个hCGB的转基因线虫株系,同时获得了3个作为功能对照研究的增强型绿色荧光蛋白(EGFP)转基因线虫株系,为深入开展胞红蛋白的功能研究奠定了基础。Objective：To investigate the function of human cytoglobin （hCGB） and to construct the transgenic Cae- norhabditis elegans expressing hCGB. Methods：The coding region of hCGB obtained from human embryo brain mRNA with RT-PCR was subcloned into the pFX-unc-ll9 to prepare the vector pFX-unc-119：：hCGB. The vectors pFX-unc-I19：：hCGB and pFX-myo-2：：dsRed were microinjected into the gonad to generate extrachromosomal strains. Integral lines were obtained by ultraviolet （UV） irradiation. Expression of hCGB in transgenic strains was detected with Western blotting and immuno- histochemistry methods. Results and Conclusion：The hCGB expressing vector was successfully constructed. Three hCGB and three EGFP transgenic strains were obtained, which will facilitate further function studies of CGB.

关 键 词：胞红蛋白 线虫纲 秀丽线虫 转基因株系 染色体 绿色荧光蛋白质类 

分 类 号：Q78[生物学—分子生物学] Q512.7