摘要
经鉴定成功构建了hPdx1和△-hPdx1蛋白表达系统,IPTG诱导蛋白表达后Western blot检测在37.2-52.3 kD之间出现了与两种目的蛋白理论分子量相符的蛋白条带。诱导后破碎菌体上清液,经分离纯化最终得到的目的蛋白纯度可达到95%以上。纯化的蛋白加入人胚胎干细胞培养基中1 h后通过细胞免疫荧光检测发现,hPdx1蛋白可进入细胞而△-hPdx1不能进入细胞,表明表达得到的蛋白具有生物活性。
hPdxl and △-hPdxl protein expression systems were identified and successfully constructed. The proteins expression were induced by IPTG and detected by Western blot, the result showed that the molecular weight of the proteins were between 37.2-52.3 kD which agreed with theories. The target proteins in the supernatant were purified and the purity could get more than 95%. The purified proteins were added into the human stem cell culture medium, respectively. The proteins were detected by the cell immunofluorescence after 1 hour, the result showed that the hPdxl protein could enter the ceils but the △-hPdx 1 can＇t, the proteins had bioactivity.
出处
《生物技术通报》
CAS
CSCD
北大核心
2012年第9期120-124,共5页
Biotechnology Bulletin
基金
国家自然科学基金项目(81070614)
