Construction of Gateway-based Destination Vectors for Detecting Subcellular

抄録

We have constructed 3 kinds of Gateway-based vectors applicable for the subcellular localization analysis of proteins in cultured Bombyx mori cells. Two were designed to visualize the location of an N- or C-terminal fusion protein with EGFP, expressed under the control of the baculoviral IE2 promoter. Another could express an N-terminal fusion protein with DsRed, similarly controlled by IE2. In order to test the applicability, the vectors were individually modified to have a cDNA encoding B. mori Actin1, histone H3 or TMS-1 (ubiquitous family of membrane protein), as well as firefly luciferase. As expected, these proteins showed cytosolic, nuclear, plasma membranous and putative peroxisomal distribution, respectively.