Hybrid transcription system for controlled plastid transgene expression

Abstract

Plastid transformation technologies have developed rapidly over the last few years, reflecting their value in the study of the principal mechanisms of plastid gene expression and commercial interest in using plastids as bioreactors. Application of this technology is still limited by the difficulty of obtaining regulated, selective expression of plastid transgenes. The plastid genome is transcribed by two different types of RNA polymerase. One of them is of the eubacterial type of polymerase, and its subunits are encoded in the plastid genome [plastid-encoded RNA polymerase (PEP)]. The other one is of the phage type and nucleus-encoded [nucleus-encoded RNA polymerase (NEP)]. To obtain selective transgene expression, we have made use of the similarities and differences between the eubacterial and the plastid eubacterial type transcription systems. We created a hybrid transcription system in which the transgene is placed under the control of a eubacterial promoter which does not exist in the plastid genome and which is not recognized by the plastid endogenous transcriptional machinery. Selective transcription of the transgene is achieved by the supply of a chimeric transcription factor that interacts with PEP and directs it specifically to the foreign eubacterial-type transgene promoter. This hybrid transcription system could be used for biotechnological and fundamental research applications as well as in the characterization of the evolutionary differences between the eubacterial and the plastid eubacterial-type transcription systems.