Abstract: CBL is a kind of Ca²⁺ sensor, which plays a pivotal role in adaptation or resistance to stress in plants. A CBL1 homologous gene was cloned from the tobacco cultivar K326, which contained a 642 bp ORF encoding 213 amino acid. The predictedmolecular weight was 24.5 kDa and the isoelectric point(pI) was 5.03. By the homology analysis, the gene had high homology with calcineurin B-like protein 1 of the Nicotiana sylvestris(NsCBL1) and Arabidopsis thaliana(AtCBL1), so named as NtCBL1. By bioinformatics analysis, NtCBL1 had the conserved EF-hand calcium binding domain of the CBL family. Expression patterns showed that the gene was expressed in roots, stems, leaves and flowers in mature stage, and the highest expression level in roots.Expression patterns under adversity stress indicated that the gene expression was induced by low-potassium, high-salt,anddrought, ABA and low-temperature, and participated in the response to biological and abiotic stress in Nicotiana tabacum. The research constructed NtCBL1-pBI121 overexpression vector.The results will provide some basis for the function analysis of NtCBL1 in response to stress.