Gene cloning of human soluble CD14 and its expression in eucaryotic cells / 中华创伤杂志(英文版)

ABSTRACT

<p><b>OBJECTIVE</b>To express human soluble CD14 (sCD14) in eukaryotic cells.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>Human sCD14 was expressed in COS-7 cells, which laid a foundation for further study.</p>