title: [Human CD96 gene cloning, expression and identification].
abstract: OBJECTIVE
  To construct and express human CD96 gene outer membrane domain (hCD96om) in prokaryotic cells and prepare rabbit polyclonal antibody of hCD96om.
  
  
  METHODS
  hCD96om was amplified by RT-PCR from the peripheral blood of patients with acute myeloid leukemia and inserted into prokaryotic expression vector pET32a(+) to construct the recombinant plasmid pET32-CD96. The expression of hCD96om was induced by IPTG in BL21(DE3) cells, and the expression product was identified by Western blotting. The anti-hCD96 polyclonal antibody was prepared by immunization of rabbits with the fusion protein. The specificity of anti-hCD96 antibody was determined by Western blotting.
  
  
  RESULTS
  hCD96om protein was expressed in E.coli BL21(DE3) cells in the form of inclusion body, with a relative molecular mass around 37 kD. Western blotting showed a specific reaction of the prepared antiserum with the 70 kD protein extracted from human leukemia cell line HL-60 cells and with the 37 kD hCD96om fusion protein.
  
  
  CONCLUSIONS
  The CD96 gene of human has been successfully cloned and expressed in BL21(DE3) cells, and its rabbit polyclonal antibody has been obtained.
authors.affiliation: Department of Laboratory Medicine, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou 510120, China.
authors.forename: Jian-ming
authors.initials: JM
authors.lastname: Zeng
authors.forename: Fei
authors.initials: F
authors.lastname: Liu
authors.forename: Ping-hai
authors.initials: PH
authors.lastname: Tan
authors.forename: Li-na
authors.initials: LN
authors.lastname: Wang
authors.forename: Mo
authors.initials: M
authors.lastname: Li
authors.forename: Zhong-hua
authors.initials: ZH
authors.lastname: Chen
authors.forename: Song
authors.initials: S
authors.lastname: Li
authors.forename: Yi-fei
authors.initials: YF
authors.lastname: Long
authors.forename: You-qiang
authors.initials: YQ
authors.lastname: Li
authors.forename: Cha
authors.initials: C
authors.lastname: Chen
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000378:metabolism; D015703:Antigens, CD / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007106:Immune Sera / Q000096:biosynthesis*; D007114:Immunization; D015470:Leukemia, Myeloid, Acute / Q000276:immunology*; D008969:Molecular Sequence Data; D014411:Neoplastic Stem Cells / Q000276:immunology; D011817:Rabbits; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: C095004:ADGRE5 protein, human; D000906:Antibodies; D015703:Antigens, CD; D007106:Immune Sera; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins