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Gene cloning of human soluble CD14 and its expression in eucaryotic cells
1Beijing Institute of Microbiology, 20, Dongdajie, Fengtai, Beijing 100071, China. yinj@nic.bmi.ac.cn
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.