Amplification and Cloning of p41-3 Gene of Plasmodium Falciparum Isolate FCC1/HN

Objective To construct a eukaryotic expression plasmid contained a gene encoding a 41 -kilodaltion blood stage antigen (p41-3) of Plasmodiu falciparum isolate FCC1 /HN. Methods According to the known sequence of p4l - 3 gene, two pairs of primers were designed and synthysized, Using PCR technique, the p4l - 3 gene was obtained by amplification from genomic DNA of isolate FCC1/HN. By cloning target gene into a eukaryotic expression vector, pcDNA3 , a recombinant plasmid pcDNA3 - p4l - 3 was constructed and transferred into E. coli DH5α. The positive recombinant pcDNA3 - p41 - 3 was screened and identified by agarose gel electrophoresis, endonuclease digestion and PCR technique. Result The p41 - 3 gene was amplified from genomic DNA of Plasmodium falciparum isolate FCC1/HN. The correct recombinant plasmid pcDNA3 - p4l -3 was identified by endonuclease digestion and PC1I technique. Conclusion The recombinant plasmid pcDNA3 -p4l - 3 was successfully constructed,which laid a foundation for expression of p41 - 3 gene in eukaryotic cell and study on the function of p41 - 3 gene.