Abstract
Approximately 10% of genes coded on the plant genome are estimated as potentially chloroplast transport proteins. However all proteins transported into the chloroplast have not been known yet. We developed cell-free translational system that worked more than 14 days. Translation is one of the most popular events in cytoplasm, and then we have an idea as development of genome-wide approach for the protein analysis based on the cell-free system. Ninety-six genes in RAFL clones with highly ChloroP score were selected and generated DNA templates by the Split-primer PCR. After transcription, these proteins were synthesized in the cell-free system and used to transport assay with isolated pea chloroplasts. Twenty-two of 42 synthesized proteins were confirmed transport into chloroplast. This indicates that this approach is suitable for genome-wide analysis of chloroplast transport proteins. As next step, phosphorylation on transit peptide (TP) of the transported proteins is investigated.
