利用SUMO融合系统高效表达可溶性重组蛋白的研究  

Efficient expression of several recombinant proteins by pSUMO expression vector

作　　者：姜媛媛[1] 尹成凯[1] 李晋南[1] 任桂萍[1] 张薇[1] 李德山[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《东北农业大学学报》2008年第10期57-62,共6页Journal of Northeast Agricultural University

基　　金：黑龙江省科技厅重点攻关项目(2006G0461-00)

摘　　要：利用pSUMO表达载体在大肠杆菌中高效可溶性表达若干种外源蛋白,如鼠源成纤维细胞生长因子(FGF)-21、人源FGF-21、人源白细胞介素(IL)-1β,并与pET-30a(+)及pTYB11表达系统作比较。将鼠源FGF-21、人源FGF-21及人源IL-1β基因分别亚克隆至pSUMO表达载体上,在大肠杆菌Rosetta(DE3)中诱导表达,并用镍离子螯合柱(Ni-NTA)纯化重组蛋白,透析后利用SUMO蛋白酶I切割融合蛋白,获得纯度较高的成熟蛋白。上述基因在pET-30a(+)及pTYB11中表达量极低,考马斯亮蓝染色几乎检测不到。在pSUMO表达体系中这些基因均以可溶形式表达,且可被SUMO蛋白酶I有效的切割,获得纯度较高的成熟蛋白。上述蛋白可在pSUMO表达系统中获得高效可溶性表达。The objective of the paper is to utilize pSUMO expression vector to efficiently express several recombinant proteins in E. coli system, including mouse fibroblast growth factor （FGF）-21, human FGF-21 and human interleukin （IL）-1β, in comparison with pET-30a（＋） and pTYBll expression systems. The genes of mouse FGF-21, human FGF-21 and human interleukin （IL）-1β were subcloned into the pSUMO expression vector. The recombinant plasmids were transformed into Escherichia coli Rosetta （DE3） and the expressed recombinant proteins were purified by His binding column. The purified recombinant proteins were dialyzed against PBS. The fusion proteins could be efficiently cleaved by SUMO protease-Ⅰ to obtain pure and active recombinant proteins. Expression level of these genes was very low in pET-30a（＋） and pTYBll expression systems and could not be detected by Coomassie blue staining. However, all these cDNAs were expressed in soluble form and the recombinant proteins accounted for 30%-50% of total bacterial proteins in the SUMO expression system. The pSUMO expression vector can efficiently express soluble recombinant proteins.

关 键 词：SUMO SUMO蛋白酶 分子伴侣 pTYB11 pET-30a(+) 表达 

分 类 号：Q786[生物学—分子生物学]