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Characterizing standard genetic parts and establishing common principles for engineering legume and cereal roots

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

DOI

  • Doreen Feike, The John Innes Centre, Norwich Research Park
    • ,
  • Andrey V Korolev, The John Innes Centre, Norwich Research Park
    • ,
  • Eleni Soumpourou, The John Innes Centre, Norwich Research Park
    • ,
  • Eiichi Murakami
    • ,
  • Dugald Reid
    • ,
  • Andrew Breakspear, The John Innes Centre, Norwich Research Park
    • ,
  • Christian Rogers, The John Innes Centre, Norwich Research Park
    • ,
  • Simona Radutoiu
  • Jens Stougaard
  • Wendy A Harwood, The John Innes Centre, Norwich Research Park
    • ,
  • Giles E D Oldroyd, The John Innes Centre, Norwich Research Park
    • ,
  • J Benjamin Miller, The John Innes Centre, Norwich Research Park, University of East Anglia

Plant synthetic biology and cereal engineering depend on the controlled expression of transgenes of interest. Most engineering in plant species to date has relied heavily on the use of a few, well-established constitutive promoters to achieve high levels of expression; however, the levels of transgene expression can also be influenced by the use of codon optimization, intron-mediated enhancement and varying terminator sequences. Most of these alternative approaches for regulating transgene expression have only been tested in small-scale experiments, typically testing a single gene of interest. It is therefore difficult to interpret the relative importance of these approaches and to design engineering strategies that are likely to succeed in different plant species, particularly if engineering multigenic traits where the expression of each transgene needs to be precisely regulated. Here, we present data on the characterization of 46 promoters and 10 terminators in Medicago truncatula, Lotus japonicus, Nicotiana benthamiana and Hordeum vulgare, as well as the effects of codon optimization and intron-mediated enhancement on the expression of two transgenes in H. vulgare. We have identified a core set of promoters and terminators of relevance to researchers engineering novel traits in plant roots. In addition, we have shown that combining codon optimization and intron-mediated enhancement increases transgene expression and protein levels in barley. Based on our study, we recommend a core set of promoters and terminators for broad use and also propose a general set of principles and guidelines for those engineering cereal species.

OriginalsprogEngelsk
TidsskriftPlant Biotechnology Journal
Vol/bind17
Nummer12
Sider (fra-til)2234-2245
Antal sider12
ISSN1467-7644
DOI
StatusUdgivet - dec. 2019

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