To study the response mechanism of tobacco to drought stress, NtSAP5 gene of zinc finger protein in tobacco was cloned and its expression under drought stress was detected in this study. The full-length coding sequence (CDS) was 474 bp and the encoding protein contains conserved zinc finger domain. The expression of NtSAP5 was regulated by drought stress, and the expression level first increased and then decreased slowly under drought stres. The CDS sequence of NtSAP5 gene was cloned into plant overexpression vector pG2BW7 and transferred to tobacco variety K326 by leaf disc method for functional verification under drought stress. Two independent transgenic lines and one non-transgenic control line were treated by drought stress. The overexpressed lines exhibited stronger drought resistance than the control. Thus, NtSPA5 played an important role in drought stress response.