Invertase catalyzes the hydrolysis of sucrose into glucose and fructose and represents a biocatalyst of great importance in food industry. Some xerophilic fungal strains have a great potential to increase enzymatic production. In the present study, five xerophilic fungal strains (Penicillium pinophilum EH2, P. purpurogenum GH2, P. citrinum ESS, Aspergillus niger GH1 and A. fumigatus GS) previously reported as invertase producers were used for isolation and cloning of an invertase gene. A fragment of 1770 bp was isolated and cloned, which corresponding to invertase gene from A. niger GH1. On the other hand, multi-banding was observed from invertase gene from other strains and invertase gene could not be isolated. Results suggested that invertase gene and amino acid sequence exhibited 97% and 99% similarity to invertase from A. niger B60 and A. kawachii IFO 4308, respectively. Therefore, A. niger GH1 could be a good alternative for invertase production.