[Construction and expression of an eukaryotic recombinant plasmid from a hybrid gene of antigen for Plasmodium falciparum].
OBJECTIVE
To construct an eukaryotic recombinant plasmid with HGFSP, a hybrid gene encoding the antigen epitopes of MSA1, MSA2, RESA and CSP in different developing stages of Plasmodium falciparum (P. f.).


METHODS
HGFSP was sub-cloned into an eukaryotic expression plasmid pcDNA3 from a prokaryotic recombinant plasmid pSK-HGFSP to construct the eukaryotic recombinant expression plasmid pc-HGFSP. The identified recombinant was then transfected into HepG2 cells with liposome-mediated method. The G418-selected positive cell clones were tested to identify the immunogenicity of HGFSP-expressing antigen.


RESULTS
It was evidenced that HGFSP was correctly inserted into pcDNA3 by restriction enzymes map analysis. HGFSP-expressing antigen-specific fluorescent response was observed in pc-HGFSP-transfected HepG2 cells. The results of SDS-PAGE and Western-blotting showed that there was a 23 kD protein band, which can be specifically recognized by anti-sera of HGFSP-expressing protein in pc-HGFSP-transfected HepG2 cell lysis.


CONCLUSIONS
Pc-HGFSP, an eukaryotic recombinant plasmid encoding hybrid antigen epitopes of P. f., was constructed successfully and the antigenicity of pc-HGFSP-expressing protein was confirmed.
Yao
Lang
L
Institute of Tropical Medicine, First Military Medical University, Guangzhou 510515. yao_lang@sina.com
Li
Ying-Jie
YJ
Li
Ming
M
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics* / Q000276:immunology; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000096:biosynthesis / Q000235:genetics*; D005057:Eukaryotic Cells / Q000378:metabolism; D006801:Humans; D010957:Plasmids / Q000096:biosynthesis / Q000235:genetics*; D010963:Plasmodium falciparum / Q000235:genetics / Q000276:immunology*; D014162:Transfection; D019444:Vaccines, DNA
D000953:Antigens, Protozoan; D004274:DNA, Recombinant; D019444:Vaccines, DNA