›› 2000, Vol. 18 ›› Issue (6): 7-346.

• 论著 • Previous Articles     Next Articles

AMPLIFICATION, CLONING AND EXPRESSION OF A GENE ENCODING HEXOSE TRANSPORTER OF PLASMODIUM FALCIPARUM

WEI Quan-de 1;YU Xin-bing 2;YE Ling 2;XU Jin 2
  

  1. 1 Department of Parasitology;Guangdong Medical College;Zhanjiang 524023; 2 Department of Parasitology;Sun Yat-sen University of Medical Sciences;Guangzhou 510089
  • Received:1900-01-01 Revised:1900-01-01 Online:2000-12-30 Published:2000-12-30

Abstract:  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 was 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.

Key words: Plasmodium falciparum, hexose transporter, gene of PfHT1, clone, expression