Abstarct: Bioremediation and phytoremediation using microorganisms are considered as advanced method to eliminate environmental contaminations. In order to identify tolerant fungi against copper contamination and study of the related genes exxpression, sampling was carried out from the soil and waste water of “Sarcheshmeh Copper Mine”, which is one of the biggest open cast copper mines in the world. Totally 73 fungal isolates were obtained and purified. Afterwards, inhibitory effect of different concentrations of CuSO4 on mycelial growth was evaluated. Results showed that concentrations of 1000, 1500, 3500 and 4500 ppm had no inhibitory effect on fungal isolates, as all of them could grow on media culture as well, only 5500 ppm of CuSO4 inhibited fungal growth compared to control. baxsed on bioassay experiments, three isolates including S3-1, S3-21 and S1-7, which were able to growth on solid and broth media containing 5500 ppm of CuSO4, were selected and identified using molecular approaches. Also, laccase and mextallothionein gene exxpression has been assessed in the selected isolates. According to molecular identification using ITS1,5.8s, ITS2 regions, isolates S3-1 and S1-7 were belonged to Pleurotus eryngii and isolate S3-21 was belonged to genus Sarocladium sp. In addition, Isolates S3-1 and S1-7 showed similar laccase gene exxpression trend, which it has been decreased after 8 days treatment with CuSO4 compared to 6th day. But, the isolate S3-21 has represented different pattern and CuSO4 treatment has increased (almost 2 times) laccase gene exxpression from 6 to 8 days. Besides, mextallothionein gene exxpression has increased from 6 days to 8 days treatment with CuSO4 compared to control which represents its role in fungal tolerance against copper in all studied isolates. In this study, Pleurotus eryngii species and genus Sarocladium sp. are introduced as heavy mextal tolerant fungi and the related gene exxpression against copper was studied for the first time in Iran