Subcloning and Expression of Two Conserved Regions (Ⅰ, Ⅴ) on P190 Antigen of Plasmodium falciparum in E. coli / 第二军医大学学报

ABSTRACT

Two DNA fragments, designated as P190CRI (AA1-55) and P190CRV (AA1597-1667) respectively, which encoded amino acid residues of conserved region I and V on the P190 antigen, were amplified by polymerase chain reaction from genomic DNA in FCC1/HN strain of Plasmodium falcipamm isolated from Hainan Province, China. It was found that there were five bases substitution in the P190CRV, in comparison with the nucleotide sequences of MAD20 strain. These two sequenced fragments were inserted into pGEX-2T plasmid. E.coli JM109 (DE3) were transformed with the recombinant plasmids and the parental plasmid. The results show that the two fragments were expressed at high level as C-terminal fusions with glutathione s-transferase (GST). The fusion proteins were easily purified from bacterial lysates by affinity chromatography using glutathione sepharose 4B.