Objective: To express human soluble CD14 (sCD14) in eukaryotic cells.Methods: Human sCD14 cDNA was amplified from U937 cells with RT PCR method. The recombinant expression plasmid pEF1/HisC/sCD14 348aa was constructed...Objective: To express human soluble CD14 (sCD14) in eukaryotic cells.Methods: Human sCD14 cDNA was amplified from U937 cells with RT PCR method. The recombinant expression plasmid pEF1/HisC/sCD14 348aa was constructed and the expression in COS 7 cells was carried out using liposome transfection method. The yield was examined with scanning map identification. The expressed product was purified by immuno affinity chromatography. Results: Sequence analysis demonstrated that the amplified gene sequence and those reported by documents were completely identical. sCD14 was expressed with high yield. The expressed product was purified to above 90%. Recombinant sCD14, specifically combinable with endotoxins, had a natural biological activity.Conclusions: Human sCD14 was expressed in COS 7 cells, which laid a foundation for further study.展开更多
基金agrantfromtheNationalNatureScienceFoundationofChina (No .394 70 76 1)
文摘Objective: To express human soluble CD14 (sCD14) in eukaryotic cells.Methods: Human sCD14 cDNA was amplified from U937 cells with RT PCR method. The recombinant expression plasmid pEF1/HisC/sCD14 348aa was constructed and the expression in COS 7 cells was carried out using liposome transfection method. The yield was examined with scanning map identification. The expressed product was purified by immuno affinity chromatography. Results: Sequence analysis demonstrated that the amplified gene sequence and those reported by documents were completely identical. sCD14 was expressed with high yield. The expressed product was purified to above 90%. Recombinant sCD14, specifically combinable with endotoxins, had a natural biological activity.Conclusions: Human sCD14 was expressed in COS 7 cells, which laid a foundation for further study.
