[Amplification, cloning and expression of a gene encoding hexose transporter of Plasmodium falciparum].
OBJECTIVE
To amplify, clone and express of a gene encoding hexose transporter of Plasmodium falcipuram (PfHT1) from Southern China isolate FCC1/HN for studing the immune of recombinant which protective from malaria parasite infection.


METHODS
Cultivation of P. falciparum isolate FCC1/HN in vitro; extraction of genomic DNA from FCC1/HN using the alkali specific cleavage method; PCR amplification of PfHT1 and cloning into eukaryotic expression vector, pEGFPN3. The recombinant as introduced into mammalian cells, HEPG2 by using liposome-mediated transfection.


RESULTS
The gene encoding PfHT1 was specifically amplified from the genomic DNA of P. falciparum isolate FCC1/HN. The size of amplified fragment was 1,516 base pair. The eukaryotic expression recombinant, pN3-HT1, was constructed and expressed steadily in the hepatocarcinoma cell lines, HEPG2.


CONCLUSIONS
The gene encoding PfHT1 was successfully amplified and cloned. The pN3-HT1/HEPG2 cell line was built for expressing fusion protein of GFP-HT1.
Wei
Q D
QD
Department of Parasitology, Guangdong Medical College, Zhanjiang 524023.
Yu
X B
XB
Ye
L
L
Xu
J
J
D000818:Animals; D003001:Cloning, Molecular; D005784:Gene Amplification; D015870:Gene Expression; D006801:Humans; D009004:Monosaccharide Transport Proteins / Q000096:biosynthesis / Q000235:genetics*; D010963:Plasmodium falciparum / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins