Transgenic crops are propagated to get a range of benefits like the ability to be resistant to fungi, bacteria, viruses and insects, to tolerate herbicides and to grow better under restricting environmental conditions such as drought, high salt or metal containing soils. Currently, GHSP-26 gene related to small heat shock protein HSP family (GHSP-26) was isolated from Gossypium arboretum and transgenic plants of Gossypium hirsutum were raised with inherited gene. Thus, the current study was carried out to evaluate the response of transgenic and non-transgenic plants (control) when they were subjected to 5 days stress by preventing the irrigation. Their response was characterized by assessing the morphological, physiological and biochemical alterations occurring in plants mechanism. After 5 days it was observed that the stress reduced the growth rate, relative water content, cell membrane stability, photosynthetic rate, transpiration rate, stomata conductance and chlorophyll in plants. The accumulation of proline and malondialdehyde took place in plants. The transgenic plants exhibited more tolerance as compared to non-transgenic plants towards the drought stress conditions and have better survival capacity in harsh environment due to the capability of diminishing their metabolic processes which can be retained when the conditions turn to normal. This relative study among the transgenic along with non-transgenic plants gives a better understanding of the transgene expression and its effect on different yield contributing traits, in this way, transgenic crops can be developed in future which will give better response under drought conditions.