Abstract: To clone the gene of human NKp46，express and purify the recombinant human NKp46 in E.coli. Methods：A hNKp46 DNA fragment，with a length of about 900?bp，was amplified from the total RNA of peripheral blood mononuclear cells by RTPCR and cloned to plasmid pMD18T，and then the cloned DNA fragment was sequenced.then hNKp46 fragment was isolated and inserted to the corresponding restriction site on procaryotic expression vector pET30a(+). The recombinant plasmid pET30a(+)hNKp46 was identified by enzymogram and transformed to E.coli BL21(DE3)，and then its expression was induced by IPTG. The expressed product was identified by SDSPAGE and Western Blotting, and the expressed protein was purified by His·Bind Purification Kit. Results：The length of DNA fragment amplified by RTPCR was consistent with that of hNKp46 cDNA. DNA sequencing of pMD18ThNKp46 revealed that the cloned DNA sequence was identical to that of reported hNKp46 cDNA. SDSPAGE proved that expressed product， with a relative molecular weight of 38.5?kD，contained about 40% of total somatic protein. Western Blotting showed that the recombinant protein could specifically bind to antiHis·Tag antibody. The recombinant protein was obtained by purification with 95.5% final purity and 40% recovery rate. Conclusion：A recombinant bacterial strain for expressing hNKp46 is successfully constructed, and its recombinant protein is purified.

Key words: NKp46, Gene clone, Prokaryotic expression, E.coli, Protein purificationNK