Gene cloning of human soluble CD14 and its expression in eucaryotic cells.
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.
Yin
Jun
J
Beijing Institute of Microbiology, 20, Dongdajie, Fengtai, Beijing 100071, China. yinj@nic.bmi.ac.cn
Bai
Jie
J
Wang
Wei
W
Song
Wei
W
Wang
Zhongze
Z
D000818:Animals; D019556:COS Cells / Q000378:metabolism; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D005057:Eukaryotic Cells / Q000378:metabolism*; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020298:U937 Cells / Q000378:metabolism
D018950:Lipopolysaccharide Receptors