The main objective of this work was to evaluate the composition, nutritional, physical and rheological properties of wheat flour and dough from genetically modified wheat (Triticum aestivum L.) Hi-Line 111 (GMW) compared to conventional wheat (non-GMW). Analyses were conducted to measure the proximate chemical composition with references to 18 components including total solid, protein, lipids, crude fiber, ash, carbohydrate, minerals, amino acids, and fatty acids. In addition, physical and rheological properties such as water absorption, arrival time, dough development time, stability value, dough weakening value, extensibility of dough, resistance to extension, and ratio of resistance/extensibility were evaluated. The results showed that there were no significant differences between GMW and non-GMW in terms of chemical composition. Results revealed the presence of saturated and unsaturated fatty acids wherein there were no significant differences between GMW and its counterpart in the levels of fatty acids. In addition, there were no significant differences on the levels of amino acids. In addition, there were no significant differences between the GMW and non-GMW in the physical and rheological properties. From these results, it can be concluded that GMW Hi-Line 111 is confirmed to have nearly the composition and rheological properties as non-GMW.