title: [Eukaryotic expression of human LIGHT-Fc chimeric molecule and its anti-tumor activity].
abstract: OBJECTIVE
  Construct eukaryotic expression plasmid of hLIGHT-Fc gene and express hLIGHT-Fc fusion protein with high biological activity.
  
  
  METHODS
  Utilizing PCR to clone the human LIGHT cDNA from a normal human activated T cells cDNA library phAD. CAD, then using SOE (splicing by overlap extension) technique to fuse CD137 signal peptide gene into LIGHT extramembrane domain encoding sequence, inserting this recombinated LIGHT cDNA with human IgGI Fc cDNA into the eukaryotic expression plasmid pcDNA3, expressed it transiently in 293T cells, and purified by recombinated protein A affinity chromatography column, at last we checked its Mr, purity and anti-tumor activity.
  
  
  RESULTS
  LIGHT-Fc gene's ORF is right and coincident with we expected, then its expression was confirmed by ELISA and SDS-PAGE, and we proved that the purified LIGHT-Fc protein could inhibit the growth of some cultured tumor cell lines by MTT.
  
  
  CONCLUSIONS
  we got a purified recombinated protein of LIGHT-Fc with anti-tumor activity and the anticipated Mr, thus lay a foundation for further works on LIGHT such as its role in immune homeostasis,the mechanism of its receptors' signal transduction, et al.
authors.affiliation: Department of Molecular Genetics, Third Military Medical University, Chongqing 400038, China. immuno02@mail.tmmu.com.cn
authors.forename: Gang
authors.initials: G
authors.lastname: Huang
authors.forename: Yun
authors.initials: Y
authors.lastname: Bai
authors.forename: Man
authors.initials: M
authors.lastname: Jiang
authors.forename: Wan-ling
authors.initials: WL
authors.lastname: Li
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics* / Q000494:pharmacology; D008565:Membrane Proteins / Q000235:genetics* / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology*; D053326:Tumor Necrosis Factor Ligand Superfamily Member 14; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D007141:Immunoglobulin Fc Fragments; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; C505914:TNFSF14 protein, human; D053326:Tumor Necrosis Factor Ligand Superfamily Member 14; D014409:Tumor Necrosis Factor-alpha