Objective To establish a mouse model of experimental autoimmune myasthenia gravis (EAMG) byimmunization with purified AchR extracted from the Narcine maculata electric organ, which meets the requirements ofinternational criteria of preclinical experiment. Methods Purified AchR protein was extracted from the electric organ ofNarcine maculata and validated by SDS-polyacrylamide gel electrophoresis. C57BL/6 mice were immunized with the proteinand complete Freund adjuvant (CFA) for the EAMG group or CFA alone for the control group three times at days 1, 30,and 60. After 70 days, the clinical assessments were determined according to body weight, serum anti-AchR antibody level,neostigmine text, and electromyography. Results Compared with control mice, the clinical scores of EAMG mice wereincreased significantly ( P < 0. 01). Body weight was significantly decreased ( P < 0. 01), neostigmine test was positive,and the percentage of decrement of the fifth wave of electromyography and levels of anti-AchR antibody were significantlyhigher ( P < 0. 01). Conclusions The C57BL/6 EAMG mouse model is successfully established by immunization withpurified AchR protein extracted from the Narcine maculata electric organ, which may provide a better research tool for studies of experimental autoimmune myasthenia gravis.