title: Gene cloning of human soluble CD14 and its expression in eucaryotic cells.
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.
authors.affiliation: Beijing Institute of Microbiology, 20, Dongdajie, Fengtai, Beijing 100071, China. yinj@nic.bmi.ac.cn
authors.forename: Jun
authors.initials: J
authors.lastname: Yin
authors.forename: Jie
authors.initials: J
authors.lastname: Bai
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Song
authors.forename: Zhongze
authors.initials: Z
authors.lastname: Wang
mesh_terms: 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
chemical_list: D018950:Lipopolysaccharide Receptors