Two field experiments were carried out at the Experimental farm of Gemmeiza Agricultural Research Station, Agricultural Research Center, Egypt, during three successive seasons; 2011/2012, 2012/ 2013 and 2013/2014. Four wheat genotypes (Triticum aestivum L.)were used in this study namely, Gemmeiza 9, Gemmeiza 11, Misr 1 and Gemmeiza line 22, which represents a wide range of drought tolerance variability and crossed to obtain F1 seeds of two crosses (Gemmeiza 9 × Misr 1) and (Gemmeiza 11 × Gemmeiza line 22). F1 plants were self-pollinated to produce F2 seeds and evaluated in two experiments. The first experiment (normal conditions) was irrigated four times after planting irrigation, the second experiment (drought conditions) was given one surface-irrigation, 30 days after planting irrigation. Presence of genes responsible for drought tolerance is basic requirement for improving any crop species including wheat. The objective of this study was conducted to assess genetic studies among two populations of bread wheat genotypes using SSR markers and SDS-PAGE aiming to developing wheat cultivars and achieving sustainability in wheat production in Egypt. The results showed that the crossing between Gemmeiza 11 and Gemmeiza line 22 which having drought tolerance in addition to good gluten strength can be used in breeding programs in future. the electrophoretic profiles of the studied genotypes of both wheat crosses revealed that the total number of protein banding patterns was twenty seven. These bands were widely distributed among wheat genotypes, having a wide range of molecular weights ranging from 16 to 127 KD. Meanwhile, band 27 in the first cross and band 10 in the second cross were unique bands characterizing the parents Gemmeiza 9 and Gemmeiza 11 and serving as markers for drought breeding.