Building the Plant SynBio Toolbox through Combinatorial Analysis of DNA Regulatory Elements
Alexander C. Pfotenhauer, Alessandro Occhialini, Mary-Anne Nguyen, Helen Scott, Lezlee Dice, Stacee A. Harbison, Li Li, Dirk Reuter, Tayler M. Schimel, C. Neal Stewart, Jacob Beal, Scott C. Lenaghan
Abstract
leaves and expression of a fluorescent reporter was measured from plant canopies, leaves, and protoplasts isolated from transfected plants. As anticipated, a dynamic level of expression was achieved from the library, ranging from near undetectable for the weakest cassette to a ∼200-fold increase for the strongest. Analysis of expression levels in plant canopies, individual leaves, and protoplasts were correlated, indicating that any of the methods could be used to evaluate regulatory elements in plants. Through this effort, a well-curated 37-member part library of plant regulatory elements was characterized, providing the necessary data to standardize construct design for precision metabolic engineering in plants.