Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag da...Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag database (EST). At the same time, it played a role in the lymphocytes differentiation and proliferation, but many of its characteristics and mechanism remain unknown up to now. To investigate its biological functions and related mechanisms, the coding region of human soluble TRAIL gene from LPS activated healthy human peripheral lymphocytes (PBL) was cloned using RT PCR and constructed a 6 His tag vector pProEX HTb hTRAIL. By DNA sequencing, the cloned hTRAIL was identical to that reported.Upon IPTG inducing, the soluble hTRAIL was highly expressed in E. coli K802 up to 30% of bacterial protein. The expressed 6 His hTRAIL protein could induce death of L929 cells as the same as TNF α. The recombinant TRAIL was obtained which paved the way for further studies on its biological characteristics and related mechanisms.展开更多
文摘Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag database (EST). At the same time, it played a role in the lymphocytes differentiation and proliferation, but many of its characteristics and mechanism remain unknown up to now. To investigate its biological functions and related mechanisms, the coding region of human soluble TRAIL gene from LPS activated healthy human peripheral lymphocytes (PBL) was cloned using RT PCR and constructed a 6 His tag vector pProEX HTb hTRAIL. By DNA sequencing, the cloned hTRAIL was identical to that reported.Upon IPTG inducing, the soluble hTRAIL was highly expressed in E. coli K802 up to 30% of bacterial protein. The expressed 6 His hTRAIL protein could induce death of L929 cells as the same as TNF α. The recombinant TRAIL was obtained which paved the way for further studies on its biological characteristics and related mechanisms.
