Random GFP::cDNA fusions enable visualization of subcellular structures in cells of Arabidopsis at a high frequency.

ABSTRACT: We describe a general approach for identifying components of subcellular structures in a multicellular organism by exploiting the ability to generate thousands of independent transformants in Arabidopsis thaliana. A library of Arabidopsis cDNAs was constructed so that the cDNAs were inserted at the 3' end of the green fluorescent protein (GFP) coding sequence. The library was introduced en masse into Arabidopsis by Agrobacterium-mediated transformation. Fluorescence imaging of 5,700 transgenic plants indicated that approximately 2% of lines expressed a fusion protein with a different subcellular distribution than that of soluble GFP. About half of the markers identified were targeted to peroxisomes or other subcellular destinations by non-native coding sequence (i.e., out-of-frame cDNAs).

SUBMITTER: Cutler SR 

PROVIDER: S-EPMC16306 | biostudies-literature | 2000 Mar

REPOSITORIES: biostudies-literature