Plastid genes in tobacco are transcribed by a plastid encoded RNA polymerase (PEP) that is related to the multisubunit RNA polymerases of eubacteria. We designed a synthetic transcription system in which promoters from the bacterium Bacillus subtilis drive the expression of a gfp reporter gene encoding a green fluorescent protein (GFP). The gfp gene is not expressed in tobacco chloroplasts, because the tobacco sigma factors do not recognize the B. subtilis promoters. The objective will be to test GFP expression from the reporter genes in chloroplasts by transient expression of engineered B. subtilis sigma factors. Transient expression studies in the summer will be followed up stable transformation of the nucleus with transgenes expressing constitutive and tissue specific chimeric sigma factors.
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