膜蛋白LASS2的表达研究  

Study of the Expression Membrane Protein LASS2

作　　者：蔡兴锋[1] 陶震[1] 曹郁[1] 杨胜利[1] 龚毅[1] 

机构地区：[1]中国科学院上海生物工程研究中心,上海200233

出　　处：《生物工程学报》2003年第1期69-73,共5页Chinese Journal of Biotechnology

摘　　要：利用多种原核表达系统、真核体外翻译系统和细菌 杆状病毒 (BactoBac)的昆虫表达系统对一个具有重要生理功能的人的膜蛋白LASS2 (Homosapienslongevityassurancehomologue 2ofyeastLAG1)进行表达研究。在原核表达系统中仅能够表达其羧基端胞外区片段却不能表达完整的LASS2蛋白 ,并制备了该片段的抗体。完整的LASS2蛋白能够在两种真核表达系统中进行表达 ,SDS PAGE分析结果表明 ,表达产物分子量为约 2 8kD的LASS蛋白 ,Western印迹分析也证实了这一结果 ,并利用Ni NTA树脂亲和层析将该蛋白纯化 ,纯度达到 90 %以上。A human membrane protein LASS2 ( Homo sapiens longevity assurance homologue 2 of yeast LAG1 ), which has important physiologic functions, was expressed in three different expression systems. Only the LASS2 protein carboxyl terminal hydrophilic fragment could be expressed in the prokaryote expression system and its polyclonal antibody was produced. The full length of LASS2 protein could be expressed successfully in both eukaryotic in vitro translation system and Bacuvirus expression system:Bac to Bac system. SDS PAGE analysis revealed that the molecular weight of expressed product of LASS2 was about 28 kD. The product was also proved by Western blot. This recombinant LASS2 protein was purified by metal affinity resin and the purity is above 90%.

关 键 词：膜蛋白 LASS2 表达研究 昆虫表达系统 真核体外翻译系统 亲和层析 

分 类 号：Q51[生物学—生物化学] Q786