人Lrp蛋白在细胞中的定位及LPS对其表达的影响  

Intracellular Location of Human Lrp and Effects of LPS on lrp Expression in Human Cell Lines

作　　者：宋庆贺[1] 于欣平[2] 陈苏民[1] 陈南春[1] 代忠明[1] 侯立朝[2] 

机构地区：[1]第四军医大学生物化学与分子生物学教研室 [2]西京医院麻醉科,西安710032

出　　处：《中国生物化学与分子生物学报》2006年第7期542-546,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.30471675)资助~~

摘　　要：为了对脂多糖应答基因(lrp)的功能进行深入的研究,用镍离子螯合柱(Ni-NTA)纯化后的全长Lrp蛋白免疫新西兰大白兔制备多克隆抗体并吸附去除非特异性反应成分.Western印迹表明,吸附纯化后的抗体可以与Lrp蛋白特异结合,并有较高免疫印迹滴度,为Lrp功能研究提供了重要的工具.激光共聚焦扫描荧光显微镜检测显示Lrp主要位于细胞核膜周围.Western印迹、RT-PCR以及细胞免疫组化染色结果都表明,用LPS刺激后,lrp在人HEK293和U937细胞内的表达均有明显的上升.结果提示,Lrp可能与对Lrp介导的反应有关.To examine the lrp gene function further, full-length Lrp protein was purified by Ni-NTA chelating column. Polyclonal antibody was prepared by immunizing rabbit using purified protein, thereafter the antiserum was adsorbed to remove the non-specific reaction components. Westem blot analysis showed that the purified antibody could bind with Lrp protein specifically, and had high titer with immunoblotting. Laser scanning confocal fluorescence microscope observation showed Lrp localized mainly around nucleus membrane. The results of Westem blot, RT-PCR and immunohistochemistry indicated that lrp expression in human HEK293 and U937 cells were both upgraded remarkably by the irritation of LPS, which suggests that Lrp probably correlate with LPS induced response.

关 键 词：脂多糖 人脂多糖应答基因 抗体制备 细胞内定位 

分 类 号：Q257[生物学—细胞生物学] Q78