Expression and immunocompetence identification of Plasmodium falciparum glutamate dehydrogenase.
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.
Li
Lin-Hai
LH
Department of Clinical Laboratory, Guangzhou General Hospital of Guangzhou Command, Guangzhou 510010, China.
Li
Ming
M
Wu
Ying-Song
YS
Wang
Ping
P
Tian
Ze-Wei
ZW
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
D016054:DNA, Protozoan; D011993:Recombinant Fusion Proteins; D005969:Glutamate Dehydrogenase