The calcineurin Blike (CBL)CBLinteracting protein kinase (CIPK) network plays a pivotal role in regulating the physiological as well as developmental processes in plants. In this study, we obtained a CIPK gene, TaCIPK16, from the wheat leaves infected by Puccinia striiformis f. sp. tritici (Pst) using in silico cloning and RTPCR. The fulllength cDNA of TaCIPK16 was 1 606 bp, which encoding 433 amino acids. Multisequence alignment showed that TaCIPK16 share high similarity with OsCIPK16 and AtCIPK16 in rice and Arabidopsis, respectively. Analysis of the protein domain features indicated that TaCIPK16 contained conserved regulatory domain in Cterminal and protein kinases domain in Nterminal. Subcellular localization assays revealed that TaCIPK16 displayed a localization pattern similar to that of the GFP control, indicating a cytoplasmic and nuclear localization. Yeast twohybrid assays showed that TaCIPK16 strongly interacts with TaCBL4 and TaCBL9, respectively. qRTPCR assays indicated that TaCIPK16 was induced by Pst infection and differentially expressed during incompatible and compatible interactions between wheat and Pst. Our results suggest that TaCIPK16 has a positive role in regulating wheat resistance against Pst.