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- W2356595997 abstract "Objective To construct the recombinant prokaryotic expression vector pET32a(+)-hDll1DSL,express and purify recombinant hDll1DSL(rhDll1DSL) protein,and investigate its effect on the expansion of cord blood CD34+ cells.Methods The hDll1DSL gene was cloned and the recombinant expression vector pET32a-hDll1DSL was constructed and transformed into E.coli BL21.The rhDll1DSL protein was expressed under the induction of isopropyl β-D-1-thiogalactopyranoside(IPTG) and purified by affinity chromatography with Ni-beads.The biological activity of rhDll1DSL was examined by recombination signal binding protein-J(RBP-J) reporter assay and expression of Hes1,a downstream effecter of the Notch pathway.The cord blood CD34+ cells were isolated using CD34 micro bead kit and incubated with hDll1DSL alone or in combination with stem cell factor(SCF),Flt3 ligand(FL) and thrombopoietin(TPO).The effect of rhDll1DSL on expansion of CD34+ cells was examined after 1-week of incubation.Results The recombinant expression vector pET32a-hDll1DSL was successfully constructed and the rhDll1DSL was successfully expressed as a Trx fusion protein(Trx-hDll1DSL) in E.coli BL21.The Trx-hDll1DSL with high purity was successfully obtained by affinity chromatography with Ni-beads.The biological activity of soluble rhDll1DSL was confirmed by activated RBP-J reporter assay and up-regulated expression of Hes1.The purified rhDll1DSL could stimulate the expansion of cord blood CD34+ cells profoundly in combination with SCF,FL and TPO.Conclusion The current study successfully constructed the recombinant expression vector pET32a-hDll1DSL which could express a biologically active Trx-hDll1DSL protein that is able to stimulate the expansion of cord blood CD34+ cells profoundly in combination with SCF,FL and TPO,and thus paved the way for its further application in optimizing the expanding culture system of human hematopoietic stem / progenitor cells in vitro." @default.
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- W2356595997 date "2011-01-01" @default.
- W2356595997 modified "2023-09-25" @default.
- W2356595997 title "Prokaryotic expression,purification of recombinant human Delta-like 1 protein and its effect on expansion of cord blood CD34+ cells" @default.
- W2356595997 hasPublicationYear "2011" @default.
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