Construction and application of a plant subcellular localization vector box of pCEG

ZHU Dan,WANG Xi,ZHU Yanming,CHEN Chao,LI Yong,BAI Xi,CAI Hua,JI Wei(College of Life Sciences,Northeast Agricultural University,Harbin 150030,China)

construction of vector box;subcellular localization;GFP;tobacco

GFP gene was widely applied in the transgenic plants and genes function's proving study.However,the construction of GFP fused plant expression vectors was always complicated and time consuming due to the hard choose of restriction enzyme cutting sites.Use plasmid pCAMBIA-1302 which included GFP gene as a original vector,and eliminated its own multiple clone site(MCS),and then inserted pET-32b's multiple clone site in front of GFP gene sequences,named the eventual plant GFP subcellular localization vector box pCEG.By Agrobacterium-mediated transformation method,pCEG instantaneous expressed in tobacco leaves,and GFP protein subcellular localization was observed by laser confocal microscope.The result displayed that the GFP proteins expressed in both cytoplasm and nucleus,this means the pCEG box can be used conveniently for plant genes subcellular localization analysis and provide a lot of usable restriction enzyme cutting site for plant genes fusioned with GFP,and the pCEG box is convenient for plant genes function analysis and has great use value.