A CBF3 gene was isolated from Jatropha curcas named JcCBF3. The full length of open reading frame(ORF)was 678 bp, encoding 225 amino acids. The amino acids sequence of JcCBF3 owned the characteristics of CBF protein. The amino acids sequences also had highly homologous with CBF genes of other plants, reached 79% with Manihot esculenta, and 74% with Hevea brasiliensis respectively. RT-qPCR result revealed that JcCBF3 was induced by low temperature . After 12 hours , the transcriptional level of JcCBF3 raised maximum 62 folds than the control, suggesting its drastic response to cold stress in Jatropha curcas. JcCBF3 was constructed into plant expression vector pBI121 and obtained the transgenic tobacco plants. At the low temperature treatment, free proline and superoxide dismutase in transgenic tobacco and the control were all increased, and that of transgenic tobacco were higher than that of normal tobacco plant. Overexpression of JcCBF3 in transgenic tobacco plants enhanced the expression of downstream genes containing CRT/ DRE elements. This study not only proves the cold resistance roles of JcCBF3, but also presents a candidate gene engineering for improvement of cold resistance in Jatropha curcas.