Objective Two primary enzymes GlpK and GlpO of glycerol metabolism from M.genitalium were chosen.This study intends to study their localization, enzyme activity and abilities to involve in the cytotoxicity of M.genitalium on host cell.Methods Prokaryotic expression vector pGEX6p-1/G1pK was constructured and GST-GlpK fusion protein was expressed.BALB/c mice were immuned by purified protein and purified antiserum titers were detected.