title: [The expression and application of human Fas ligand in E.coli].
abstract: OBJECTIVE
  To express recombinant human FasL molecule in E.coli.
  
  
  METHODS
  RT-PCR was applied to amplify FasL cDNA from the total RNA extracted from activated human peripheral blood lymphocytes. The DNA fragment was cloned into PCR2.1 vector. After sequencing, the FasL gene was inserted into pQE-31 vector and expressed in E.coli M15. The FasL protein was purified through Ni-ATA affinity chromatography column and identified by SDS-PAGE and Western blot. The mice were immunized with the FasL protein and the specific anti-serum was harvested 6 weeks after immunization. The serum level of FasL from with different kinds of diseases patients were detected using the anti-FasL antibodies from the immunized mice.
  
  
  RESULTS
  The expressed protein could be recognized by anti-human FasL antibody in Western-blot analysis with M(r)40 000. This protein could induce Jurket cells apoptosis. anti-FasL serum prepared from mouse could detect the serum FasL as sensitive as commercial ELISA kits.
  
  
  CONCLUSIONS
  The human FasL protein is obtained. It lays the foundation for the further detecting the concentration of FasL and sFasL of patients.
authors.affiliation: Department of Immunology, Fourth Military Medical University, Xi'an 710032, China. ningli@shsmu.edu.cn
authors.forename: Ning-li
authors.initials: NL
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Nie
authors.forename: Qi-wen
authors.initials: QW
authors.lastname: Yu
authors.forename: Jun
authors.initials: J
authors.lastname: Bo
authors.forename: An-lun
authors.initials: AL
authors.lastname: Ma
authors.forename: Ji-ying
authors.initials: JY
authors.lastname: Zhang
authors.forename: Bai-hua
authors.initials: BH
authors.lastname: Shen
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Dong-qing
authors.initials: DQ
authors.lastname: Zhang
mesh_terms: D000818:Animals; D017209:Apoptosis; D003001:Cloning, Molecular; D003922:Diabetes Mellitus, Type 1 / Q000097:blood*; D004926:Escherichia coli / Q000378:metabolism*; D053222:Fas Ligand Protein; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D019169:Jurkat Cells / Q000166:cytology; D007938:Leukemia / Q000097:blood; D008180:Lupus Erythematosus, Systemic / Q000097:blood; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013274:Stomach Neoplasms / Q000097:blood; D048069:Tumor Necrosis Factors / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: C505661:FASLG protein, human; D053222:Fas Ligand Protein; C505658:Fasl protein, mouse; D007106:Immune Sera; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D048069:Tumor Necrosis Factors