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[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.
Conclusion:
The gene encoding PfHT1 was successfully amplified and cloned. The pN3-HT1/HEPG2 cell line was built for expressing fusion protein of GFP-HT1.