Calmodulinlike protein (CML) is a very important type of calciumsensing protein in plant cells. Studies have shown that CML proteins play critical roles in pollen germination and pollen tube growth. In this study, using the selfpollinated stigma of the Chinese cabbage selfincompatibility line ‘91125’, we cloned the fulllength cDNA sequence of BrCML49 gene and obtained with 1 046 bp, including 960 bp open reading frame, encoding 319 amino acids, having 2 conserved EFhand domains. Sequence analysis found that the BrCML49 protein is a stable hydrophilic protein and does not have the transmembrane region. Realtime quantitative expression analysis (qRTPCR) showed that BrCML49 gene exhibited the highest expression in the anther of ‘91125’, followed by the whole flower and petal, and the expression level was lower in other tissues. In the Chinese cabbage selfcompatibility line ‘14S23’, the BrCML49 gene was highly expressed in the whole flower and stigma, and the expression level is lower in other tissues. Subcellular localization analysis revealed that BrCML49 was mainly localized in the cell membrane and nucleus of tobacco leaf. The results of this study can lay a foundation for further study on the function of Chinese cabbage BrCML49 gene, and provide a theoretical basis for elucidating the signaling mechanism of Chinese cabbage BrCML49 gene involved in selfincompatible pollen germination and pollen tube growth.