[Cloning and sequencing of the genes coding for the histidine-rich protein II of Plasmodium falciparum]

Abstract

Aim: To compare and analyze the homology of genes encoding histidine-rich proteinII (HRPII) of different Plasmodium falciparum isolates.

Methods: Using PCR technique, the complete genes coding for HRPII of P. falciparum isolates FCC1/HN and VN isolates were amplified. PCR products were digested by HindIII/BamHI and cloned into plasmid pUC19. The recombinant plasmid HRPII/pUC19 was screened and identified by PCR and restriction analysis. The cloned HRPII genes were sequenced by Sanger's method.

Results: HRPII genes of FCC1/HN and VN isolates were successfully amplified and cloned into pUC19. DNA sequencing showed that the coding length of HRPII gene was 1,020 bp without introns in FCC1/HN and VN isolates, however, there were ten points mutations between them. FCC1/HN isolate exhibited 98.8%, 92.2% and 98.7% homology in amino acids with isolates VN, IMTM22, and Itg2, respectively. Though the numbers of repeat sequences were different in four isolates, they had the same hydrophobic leader sequence and a single putative glycosylation site. The secondary structure analysis showed that the main antigenic determinants of four isolates were located on 5' end non-repeat region (amino acids 1-60).

Conclusion: FCC1/HN isolate was highly homologous in the coding region of HRPII with VN, IMTM22, and Itg2 isolate. Four isolates exhibited similar structural characteristics and antigenic determinants in HRPII.