title: [The expression of C-type lectin-like domain of DNGR-1 molecules of Chinese rhesus macaques in E.coli].
abstract: OBJECTIVE
  To obtain the nucleotide sequence and express the C-type lectin-like domain (CTLD) of C-type lectin domain family 9, member A (Clec9A) gene of Chinese rhesus macaques.
  
  
  METHODS
  The complete coding region of rhesus macaque Clec9A gene was amplified by sequence specific RT-PCR and sub-cloned into the pGEM-T vector and sequenced. Recombinant plasmid pET-32a-CTLD was constructed to express the CTLD region of CLEC9A in BL21 (DE3) cells under IPTG induction. The recombinant protein was identified by SDS-PAGE and Western blotting.
  
  
  RESULTS
  The 726 bp complete coding sequence of Clec9A gene was obtained. Sequencing and sequence alignments showed that rhesus macaque Clec9A gene shared 95.0% and 73.1% nucleotide identities and 91.7% and 57.3% amino acid identities with humans and mice, respectively. In vitro expression of CTLD resulted in a protein with Mr; of 32 000. Western blotting revealed that the protein had the similar antigenicity with human DNGR-1.
  
  
  CONCLUSIONS
  The Clec9A gene of Chinese rhesus macaques is highly similar to that of humans and could code a protein with the antigenicity of human CTLD protein.
authors.affiliation: National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
authors.forename: Wenrong
authors.initials: W
authors.lastname: Yao
authors.affiliation: National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
authors.forename: Guibo
authors.initials: G
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002681:China; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D037181:Lectins, C-Type / Q000235:genetics / Q000378:metabolism*; D008253:Macaca mulatta / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011975:Receptors, Mitogen / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: C530206:CLEC9a protein, human; D037181:Lectins, C-Type; D011975:Receptors, Mitogen; D011993:Recombinant Fusion Proteins