In this research, there were 41 tobacco lines with resistance to kanamycin generated from genetic
transformation. The kanamycin-tolerant plants were analyzed by PCR to screen out the positive transformants.
Finally, 24 lines contained the Pp14-3-3 gene were confirmed, and the integration of Pp14-3-3 into the
genome of the tobacco lines indicated all the 24 lines were positive transformants. A total of 3 T1 transgenic
tobacco lines, 3-5, 3-7 and 3-10, were chosen to be further studied in the following assays. In order to detect
whether the Pp14-3-3 gene was expressed under normal and stressful conditions, the total RNA was isolated
from the young leaves for real time PCR analysis. As a result, the Pp14-3-3 was highly expressed in the
transgenic lines under normal and stressful conditions. The transcriptional level of Pp14-3-3 was slightly
induced by drought and NaCl stresses. Doubtlessly, the Pp14-3-3 was overexpressed in the T1 generation
plants.