title: Expression and immunocompetence identification of Plasmodium falciparum glutamate dehydrogenase.
abstract: OBJECTIVE
  To express the fusion protein of glutamate dehydrogenase (GDH) with glutathione S-transferase (GST) of Plasmodium falciparum FCC1/HN in E. coli BL21 and assess the immunocompetence of the recombinant protein.
  
  
  METHODS
  GDH gene of P. falciparum was specifically amplified with PCR, followed by double enzyme digestion and cloning the gene fragment into pGEX-4T-1 vector for the expression of the fusion protein GST. The recombinant plasmid was transformed into E. coli BL21. Four mice (Kunming strain) were immunized with purified recombinant protein (antigen) and the polyclonal antibodies produced in response to the treatment were collected. Enzyme-linked immunosorbent assay and Western blotting were carried out to examine the immunocompetence of the recombinant protein.
  
  
  RESULTS
  The fusion protein was successfully expressed, which exhibited specific reaction with the sera obtained from mice immunized with P. falciparum. Specific humoral responses were elicited after introducing the fusion protein in mice and the specific antibody titer was 1:16 in agar diffusion assay.
  
  
  CONCLUSIONS
  GDH of P. falciparum may have successful expression in E. coli BL21 and the expressed protein possesses high antigenicity.
authors.affiliation: Department of Clinical Laboratory, Guangzhou General Hospital of Guangzhou Command, Guangzhou 510010, China.
authors.forename: Lin-Hai
authors.initials: LH
authors.lastname: Li
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.forename: Ying-Song
authors.initials: YS
authors.lastname: Wu
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.forename: Ze-Wei
authors.initials: ZW
authors.lastname: Tian
mesh_terms: D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan; D015870:Gene Expression; D005822:Genetic Vectors; D005969:Glutamate Dehydrogenase / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007121:Immunocompetence; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
chemical_list: D016054:DNA, Protozoan; D011993:Recombinant Fusion Proteins; D005969:Glutamate Dehydrogenase