Structure and expression analysis of the OsCam1-1 calmodulin gene from Oryza sativa L.
Calmodulin (CaM) proteins, members of the EF-hand family of Ca(2+)- binding proteins, represent important relays in plant calcium signals. Here, OsCam1-1 was isolated by PCR amplification from the rice genome. The gene contains an ORF of 450 base pairs with a single intron at the same position found in other plant Cam genes. A promoter region with a TATA box at position-26 was predicted and fused to a gus reporter gene, and this construct was used to produce transgenic rice by Agrobacterium-mediated transformation. GUS activity was observed in all organs examined and throughout tissues in cross-sections, but activity was strongest in the vascular bundles of leaves and the vascular cylinders of roots. To examine the properties of OsCaM1-1, the encoding cDNA was expressed in Escherichia coli. The electrophoretic mobility shift when incubated with Ca(2+) indicates that recombinant OsCaM1-1 is a functional Ca(2+)-binding protein. In addition, OsCaM1-1 bound the CaMKII target peptide confirming its likely functionality as a calmodulin.
Phean-o-pas
Srivilai
S
Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
Limpaseni
Tipaporn
T
Buaboocha
Teerapong
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D002147:Calmodulin / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D012275:Oryza / Q000033:anatomy & histology / Q000166:cytology / Q000235:genetics*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA
D002147:Calmodulin; D011994:Recombinant Proteins