Abstract: Cytosolic glyceraldehyde 3-phosphate dehydrogenase (GAPC) takes part in glycolysis and is a classical housekeeping enzyme in both prokaryotes and eukaryotes. Increasing evidence indicates that GAPC is involved in various plant abiotic and biotic stress responses such as salt,low phosphate,reactive oxygen species (ROS) and heat in phytophthora infestans. The AtGAPC2 gene has been found in responses to low phosphate and osmotic stress in Arabidopsis. Analysis of sequences and protein structure of GAPC could help in studies of the function and mechanisms of GAPC acting in plant stress resistance. The fragments encoding GAPC were obtained from Solanum tuberosum (StGAPC) and Arabidopsis thialiana (AtGAPC2) seedlings by reverse transcription polymerase chain reaction (RT-PCR). The CDS lengths of StGAPC and AtGAPC2 were 1017 bp,encoding a protein subunit of 338 amino acids. Bioinformatics analysis demonstrated that the StGAPC and AtGAPC2 had 84% similarity in nucleotide sequence and 92% similarity in amino acid sequence. Molecular weights of the proteins were 36.65 and 36.91 kDa,the PI (theoretical isoelectric point) were 6.34 and 6.67,and both of them had stable protein structures. The 3D structures of StGAPC and AtGAPC2 were predicted by homology comparative modeling in the Swiss-Model,and showed that the 3D structures were highly similar to Oryza sativa GAPDH C chain (2e5rC). Predicted proteins contained conserved GAPC domains which included an NAD binding domain constructed by βαβαβ Rossmann coil and a catalyzed domain in the saddle super secondary structure composed of seven β foldings. CDS sequences of GAPC showed that the same family of plants has a high homology attributed to the same branch in the evolutionary tree and this evolutionary relationship corresponded to the existing classification system. pBI121 was constructed as a base vector and the gene expression vector in the plant was driven by a CaMV35S promoter. The recombinant vector was introduced into Agrobacterium strain EHA105 and positive clones were determined by PCR. This research is a basis to obtain transgenic materials and for further study on the function of GAPC.