Objectives: Sleep quality is directly related to the health, quality of life and working capacity and is affected by age, gender, exercise, life habits. Gender dissimilarities in sleep quality are acknowledged. However, the gender difference in the quality of sleep in shift workers was not well known. Our aim was to study the less known gender differences in the quality of sleep of shift workers. Methods: 1,008 study subjects aged between 21 and 57 years among these, 637(525 male and 112 female workers) were daytime workers and 371(253 males and 118 female workers) were shift workers. Sleep quality was measured by self-report through the Pittsburg Sleep Quality Index(PSQI) and fatigue was measured by using the Chalder Fatigue Scale(CFS). All data was analyzed using SPSS version 19.0 program. The study was approved by the Institutional Review Board of the Occupational Safety and Health Research Institute. Results: Overall, sleep quality was found to be worse in shift workers than in daytime workers and in female than in male. Fatigue in shift female workers was significantly higher than in daytime female workers and shift male workers. In Spearman correlation analysis results, fatigue were significantly associated with gender, drink habit, regular exercise, working hours and sleep quality. With group male workers as the reference, the odds ratio(OR) for having a CFS was 2.115(${\beta}$ value=0.749, p<0.01), and subjective sleep quality, sleep latency, daytime dysfunction and PSQI were 1.541(${\beta}$ value=0.432, p<0.05), 2.297(${\beta}$ value=0.831, p<0.001), 1.798(${\beta}$ value=0.587, p<0.01) and 2.224(${\beta}$ value=0.799, p<0.01), respectively. Conclusions: Shift work played an important role in lowering the quality of sleep, and the effect was more pronounced in female workers than in male workers. Sleep quality was related to fatigue, especially sleep quality, sleep duration, sleep disturbances and daytime dysfunction were significantly related. Therefore, to improve fatigue and sleep quality of shift worker, it is necessary to improve healthy habits such as regular exercise, smoking cessation, and limited drinking. In particular, it is urgent to develop and implement a regular exercise program to reduce fatigue and improve the quality of sleep in workplace.