To investigate the amino acid sequence of tobacco Nt-GAD at the molecular level,the physical and chemical characteristics,structural domains and evolutionary trees of tobacco Nt-GAD amino acid sequence were analyzed using the tools of Protparam,Interpro,Blast,Muscle,Phyml and Treedin.The results showed that the isoelectric points of tobacco Nt-GAD members ranged between 5.5～6,and Nt-GAD1 exhibited the highest homology with Nt-GAD2.In addition,Nt-GAD2 had the best heat resistance and Nt-GAD3 had the strongest stability.Tobacco Nt-GAD was classified within the pyridoxal phosphate-dependent decarboxylase fami-ly and the glutamate decarboxylase family,possessing the pyridoxal phosphate-dependent transferases homologous superfamily domain.A distinctive calmodulin domain was located at the C-terminal.Based on evolutionary tree analysis,tobacco Nt-GAD members were categorized into three groups.Among these group,tobacco Nt-GAD3(group 1)is more prominent,whereas Nt-GAD2(group 3)is more conservative,Nt-GAD1 and Nt-GAD4 con-stitute the second group.
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