Gene cloning of human soluble CD14 and its expression in eucaryotic cells

摘要

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 immune-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.