摘要
黑腹果蝇是生命科学领域研究常用的经典模式生物。本研究以黑腹果蝇为样品,在总蛋白提取、蛋白纯化和等电聚焦条件等三个方面对果蝇蛋白质组双向电泳条件进行优化,为果蝇比较蛋白质组学研究奠定基础。结果显示:(1)蛋白提取:利用Bullet Blender细胞组织破碎仪处理样品优于超声破碎及液氮研磨;(2)蛋白纯化:利用2D-clean up试剂盒纯化蛋白效果较好;(3)等电聚焦:等电聚焦总volt-hours在15 000 volt-hours左右时竖条纹相对较少;通过综合优化后的双向电泳技术所得到的蛋白图谱中蛋白点相对较多且圆滑,条纹现象较轻,重复性较好,满足后续软件分析以及数据处理的要求,适用于黑腹果蝇以及其他蝇类的蛋白质组学研究。
Drosophila melanogaster is a classic model organism in life science research. In this study, conditions of two-dimensional electrophoresis for Brosophila melanogaster were optimized in three aspects, including protein extraction, protein purification and isoelectric focusing condition. Results showed that： （1） Protein extraction： by comparing with ultrasonic and liquid nitrogen grinding, Bullet Blender had a better effect on sample breaking; （2） Protein purification： by comparing both protein purification methods, the 2D-clean up kit had a better performance; （3） Isoelectric focusing： when the total volt-hours of isoelectric focusing was at 15 000 volt-hours, the vertical stripes were relatively less. After the comprehensive optimization of two-dimensional electrophoresis, the acquired maps had more and clearer protein spots, less stripes and better reproducibility, meanwhile, it could accord with the requirements of software analysis and experimental treatments. The optimization could be applied to the proteomic study of drosophila melanogaster and other fly species.
作者
刘清秀
侯微
王权威
闫梅霞
崔丽丽
王英平
Liu Qingxiu Hou Wei Wang Quanwei Yan Meixia Cui Lili Wang Yingping(College of Traditional Chinese Medicine, Jilin Agricultural University, Changchun, 130118 Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, 130112 Key Laboratory for Cultivation Technology of Traditional Chinese Medicines in Jinlin Province, Changchun, 130112)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2016年第9期2324-2328,共5页
Genomics and Applied Biology
基金
吉林省现代农业产业技术体系建设研究课题(特色红参超微粉系列产品开发与示范(2015026))资助
关键词
黑腹果蝇
蛋白质组
双向电泳
优化
Drosophila melanogaster, Proteome, Two-dimensional electrophoresis, Optimization
