Recent studies have found that early nodulin-like (ENODL) genes play a vital role in the formation and development of underground storage organs in plants. Our previous study found that some ENODL genes were expressed abundantly during the root formation and tuber expansion stages of Tetrastigma hemsleya-num. In this study, two ENODL genes ThENODL1 and ThENODL2 from T. hemsleyanum were cloned. Preliminary bioinformatics analysis showed that the open reading frame (ORF) for ThENODL1 is 528 bp, enco-ding 175 amino acids, and the ORF for ThENODL2 is 651 bp, encoding 217 amino acids. Both ThENODL1 and ThENODL2 have the conserved domain of rice ENODL gene (OsENODL1_like), and possess the basic molecular structure characteristics of ENODL gene. Relative fluorescence quantitation PCR revealed that the expression level of ThENODL1 was significantly higher in tuberous roots than in other organs, and signifi-cantly increased in the initial development stage and expansion stage of root tuber, while the expression level of ThENODL2 was the lowest in tuberous roots and there was no significant difference in expression profiles at different development stages of root tuber. Furthermore, the proteins encoded by cloned cDNAs of both ThENODL1 and ThENODL2 were expressed with the expected size using E. coli prokaryotic expression sys-tem. This study lays a basic molecular biology foundation for further ascertaining the functions of ThENODL genes in tuber development.