Objective To amplify the hexokinase (HK) gene of Plasmodium falciparum isolate FCC1/HN, compare the sequence with that of other P. falciparum isolates and human, and construct its prokaryotic and eukaryotic expres-sion plasmlds. Methods The HK gene was amplified by PCR from the genomic DNA of P. falciparum isolate FCC1/HN. The amplified HK gene was cloned into the prokaryotic and eukaryotic expression vectors, pET-30a(+) and pcD-NA3, and transferred into Escherichia coli BL21/DE3 and JM109, respectively. The positive recombinant pET-30a(+)-HK and pcDNAa-HK were screened and identified by endonuclease digestion and PCR. The nucleotide sequence of HK gene was determined by the dideoxy chain termination method. The DNA sequence and its deduced protein sequence were analyzed. Results The HK gene of P. falciparum isolate FCCI/HN was specifically amplified, and the correct recom- binant plasmids pET-30a(+)-HK and pcDNA3-HK were constructed. The results of sequencing the nucleotide acids showed that the full length HK gene of P. falciparum isolate FCC1/HN was 1 482 base pair. It encoded a protein of 493 amino acids. The amino acids sequence was the same as that of isolate 3D7, with 99.8% identity of the isolate K1, but with 23.2%-26.6% identity of all the 4 types of the human HK. Conclusion The HK gene of P, falciparum isolate FCCI/HN was amplified. Its recombinant prokaryotic and eukaryotic expression plasmids were successfully constructed and its nucleotide sequence were determined. The HK gene of P. falciparum is quite conserved among different isolates, but has a low homology with human HK. The HK of P. falciparum may be a potential drug target.