Abstract: RT-PCR was used to obtain two genes named as HbCBL1 and HbCBL2 from Hordeum brevisubulatum.Protein structure analysis showed that the proteins encoded by these two genes have the typical EF-hands,and HbCBL1 has N-myristoylation motif.In order to clarify whether HbCBL1 and HbCBL2 protein structures contribute to their subcellular localization,several plant expression vectors were constructed and transformed into onion epidermal cells by particle bombardment,respectively.These vectors were also transformed into Arabidopsis by floral dip.The subcellular localization of fusion proteins was determined by Confocal laser scanning microscope.The results of transient expression showed that the fusion protein HbCBL2-GFP was accumulated in tonoplast,but the HbCBL1-GFP fussion protein was accumulated in plasma membrane as well as in nucleus;N-myristoylation motif of HbCBL1 involves in its localization at cell level.The results of stable expression showed that HbCBL2 was located at tonoplast and HbCBL1 was located at plasma memebrane,additionally,both HbCBL2 and HbCBL1 are located at the guard cells.It was suggested that HbCBL1 and HbCBL2 protein structures involved in their subcellular localization,which play roles for further analyses of gene functions.

Key words: Hordeum brevisubulatum, HbCBL2, HbCBL1, Confocal laser scanning microscope ( CLSM), Subcellular localization

CLC Number: 

• Q71