1. Gene cloning of human soluble CD14 and its expression in eucaryotic cells.
- Author
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Brawarsky P, Brooks DR, Wilber N, Gertz RE Jr., Walker DK, Yin, Jun, Bai, Jie, Wang, Wei, Song, Wei, and Wang, Zhongze
- Abstract
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. [ABSTRACT FROM AUTHOR]- Published
- 2002