title: [Expression of human prp gene in prokaryotic cells using GST fusion protein expression system].
abstract: OBJECTIVE
  To study the biological features of cell-surface protein PrPc, which is thought to be involved in the prion-associated diseases after converting to a proteinase-resistant isoform PrPSc posttranslationally, and to establish an effective immunologic diagnostic method using PrPc as antigen.
  
  
  METHODS
  Amplifying and cloning the human prp gene from lymphocytes of two normal Chinese, after confirmed by DNA sequence analysis, the genes were separately subcloned into a GST-fusion expression plasmid.
  
  
  RESULTS
  Sequence analysis showed that one contatined a point mutation that induced the 65th amino acid "Trp" inverting to a stop codon "TAG", whereas the other had the same sequence as the published standard prp gene. Both the standard and the mutated prp genes were separately subcloned into a GST fusion protein expression vector and expressed in the prokaryotic cells effectively. Western blot assay revealed that both of them expressed GST-PrP fusion proteins and could be recognized by PrP specific monoclonal antibody.
  
  
  CONCLUSIONS
  It suggests that human PrP protein can be expressed in the GST fusion protein expression system and the expressed proteins hold good immune-reactivity.
authors.affiliation: Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052.
authors.forename: X
authors.initials: X
authors.lastname: Zhao
authors.forename: X
authors.initials: X
authors.lastname: Dong
authors.forename: T
authors.initials: T
authors.lastname: Hung
mesh_terms: D000595:Amino Acid Sequence; D005982:Glutathione Transferase / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D018620:PrPSc Proteins / Q000235:genetics; D017096:Prion Diseases / Q000235:genetics; D011328:Prions / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D018620:PrPSc Proteins; D011328:Prions; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase