AMPLIFICATION, CLONING AND EXPRESSION OF A GENE ENCODING HEXOSE TRANSPORTER OF PLASMODIUM FALCIPARUM
- VernacularTitle:恶性疟原虫己糖转运体编码基因的体外扩增、克隆及表达
- Author:
Quande WEI
;
Xinbing YU
;
Ling YE
;
Ji XU
- Publication Type:Journal Article
- Keywords:
Plasmodium falciparum, hexose transporter, gene of PfHT1, clone, expression
- From:
Chinese Journal of Parasitology and Parasitic Diseases
1997;0(06):-
- CountryChina
- Language:Chinese
-
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.
