Objective To clone and study the antimicrobial-resistant gene in Neisseria gonorrhoeae.Methods The gene library,which contains the differential genes of antimicrobial resistant strains and stan-dard reference strains of Neisseria gonorrhoeae,was constructed using a technique known as suppression subtractive hybridization(SSH).Then the antimicrobial resistance associated genes were cloned and ana-lyzed.Results Subtractive gene library in antimicrobial-resistant Neisseria gonorrhoeae was successfully constructed,which contains2500positive clones.Sequence analysis was performed on5clones.The se-quences of these five clones were unknown previously.Conclusions The subtractive DNA library is succes-sively constructed which may provide an important clue for studying the mechanism of antimicrobial resis-tance in Neisseria gonorrhoeae.