重组人sIL-2Rα在HEK293细胞中的分泌性表达与鉴定  

Secretory expression and identification of recombinant human sIL-2Rα in HEK293 cells

作　　者：王双[1] 陈燕侠[2] 王瑞琳[1] 孙志伟[1] 林建波[1] 俞炜源[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071 [2]空军总医院干部病房,北京100036

出　　处：《军事医学科学院院刊》2010年第5期430-432,共3页Bulletin of the Academy of Military Medical Sciences

摘　　要：目的在HEK293细胞瞬时表达带有组氨酸标签的重组人sIL-2Rα的胞外功能区片段,并进行纯化与鉴定。方法对GenBank中IL-2Rα膜外区基因序列进行优化,并在3′端融合组氨酸标签序列的基因,人工合成该基因并将其克隆到表达载体pL-293;转染HEK293细胞瞬时表达sIL-2Rα;表达产物His标签纯化;SDS-PAGE和Western印迹对表达产物进行鉴定。结果表达产物中在相对分子质量为48 000处可见与目的蛋白相对分子质量相符的条带,该条带可被sIL-2Rα单克隆抗体识别。结论构建的重组sIL-2Rα质粒能够在HEK293细胞瞬时表达,表达量为1~1.5 mg/L。Objective To express transiently the functional fragment of recombinant sIL-2Rα protein by HEK293 and to purify it after identification.Methods The genes encoding the human sIL-2Rα protein were optimized for mammalian cell expression and chemically synthesized,with a 6×His tag at C-terminal.The genes were cloned into the pL293 vector to create the recombinant plasmid pL293-sIL-2R which was transfected in human embryo kidney 293F cells.The gene expression products were identified by Western-blot and purified by His-tag.Results The expression products were about 48 000（Mr）,which could be identified by mAb of sIL-2Rα proteins.Conclusion The sIL-2Rα can be secretorily and transiently expressed from HEK293 cells and protein product yield is 1 to 1.5 mg/L cells.

关 键 词：SIL-2R αHEK293 瞬时表达 

分 类 号：Q78[生物学—分子生物学]