摘要：The eDNA encoding human TRAIL extraeellular region(amino acids 41-281) was amplified byreverse tran...The eDNA encoding human TRAIL extraeellular region(amino acids 41-281) was amplified byreverse transcription(RT)-PCR from total RNA derived from human acute promyelocytic leukemia cell line HL60. After sequencing, the cDNA was cloned into the vector pQE-80L and transformed into E.coli DH5aα.By IPTG induction, the soluble TRAIL41-281(sTRAIL41-281) protein was expressed with the 40% of total bacteria protein. Inclusion bodies were dissolved into 8 mol/L urea, purified by Ni-NTA chromatography column, the product with over 90% purity was obtained. After refolding by dialysis, the active trimer form of sTRAIL41-281 was derived from the renatured proteins by gel filtration chromatography. The MTT assay, flow cytometry and DNA fragmentation assay showed that the refolded sTRAIL41-281 could potently inhibit the growth of Jurkat cells and induce apoptosis, confirmed the apoptosis-inducing activity of sTRAIL41-281 on tumor cells, it will benefit the further research of TRAIL.显示全部
