摘要
为获得hEGF在大肠杆菌中的高效表达,构建了pPTD-hEGF原核表达载体。将其转化大肠杆菌BL21(DE3),经0.3mmol/LIPTG诱导,PTD-hEGF融合蛋白进行了有效表达,表达产物主要形成了包涵体。经Ni2+-NTA亲和层析纯化,融合蛋白的纯度达99.5%。MALDI-TOF-MS分析证明表达的融合蛋白氨基酸序列完全正确。PTD-hEGF融合蛋白能明显促进HEK-293细胞的分裂和生长。
To produce membrane-permeable human epidermal growth factor （hEGF）, a pPTD-hEGF prokaryocyte expression vector was constructed and transformed into E. coli BL 21 （DE3）. The PTD-hEGF fusion protein was induced by 0.3mmol/L of IPTG expressed in the form of inclusion body with an yield of 40 % of the total protein in the cells, and then purified by Ni^2 ＋ NTA affinity chromatography. The SDS-PAGE analysis showed a single fusion protein band with a molecular weight of 16kD. The amino acid sequence was checked by MALDI-TOF-MS analysis. The genetic engineering PTD-hEGF fusion protein obviously promoted the proliferation and growth of the HEK-293 cells in vitro.
出处
《生物工程学报》
CAS
CSCD
北大核心
2007年第4期589-592,共4页
Chinese Journal of Biotechnology
