Cloning and expression of the gene of Plasmodium falciparum FCCI/HN encoding purine nucleoside phosphorylase

Amplification, cloning and expression of the gene encoding the purine nucleoside phosphorylase (PNP) of Plasmodiun falciparum FCC1/HN, isolated from the southern China isolate. According to the published PNP gene sequence from Plasmodi-um falciparum, a pair of oligonudeotides was designed as primers, the gene of the purine nucleoside phosphorylase of the FCC1/HN isolate of Plasmodium falciparum was amplified by using PCR technique.The PCR product was purified and digested with EcoR 1 and Xho I , then cloned into the plasmid pET30a( + ) and pcDNA3 at the EcoR I and Xho I sites. The recombinant plasmid were i-dentified by restriction analysis, PCR amplification and DNA sequence analysis, then the recombinant plasmid pET30a( + )-PNP was transformed into E.coli BL21(DE3) .The recombinant pET30a( + )-PNP was induced by IPTG concentration. A high level expression of PNP fusion protein was analyzed. The results showed that the gene of PNP was amplified from the genome of Plasmodiun falciparum FCC1/HN, the southern China isolate and the purified PCR product was directly inserted into the plasmids pET30a( + )and pcDNA3 at the EcoR I and Xho I sites.The PNP fusion protein induced by IPTG concentration was analyzed by SDS-PAGE, the fusion protein molecular weight is 31.4kDa. It concludes that PNP gene is obtained from genome of Plasmodium falciparum and the recombinant plasmids of the pET30a( + )-PNP and pcDNA3-PNP are successfully constructed .The PNP fusion protein expresses in E. coli BL21(DE3) and analyzed by SDA-PAGE.