Gene cloning of human soluble CD14 and its expression in eucaryotic cells.
- Author:
Jun YIN
1
;
Jie BAI
;
Wei WANG
;
Wei SONG
;
Zhongze WANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; COS Cells; metabolism; Chromatography, Affinity; Cloning, Molecular; Eukaryotic Cells; metabolism; Humans; Lipopolysaccharide Receptors; genetics; Reverse Transcriptase Polymerase Chain Reaction; U937 Cells; metabolism
- From: Chinese Journal of Traumatology 2002;5(3):156-160
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo express human soluble CD14 (sCD14) in eukaryotic cells.
METHODSHuman 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.
RESULTSSequence 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.
CONCLUSIONSHuman sCD14 was expressed in COS-7 cells, which laid a foundation for further study.
