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A Facile Agrobacterium‑Mediated Transformation Method For The Model Unicellular Green Algae Chlamydomonas Reinhardtii, Truyen N. Quach, Shirley J. Sato, Mark R. Behrens, Paul N. Black, Concetta C. Dirusso, Heriberto D. Cerutti, Tom Elmo Clemente
A Facile Agrobacterium‑Mediated Transformation Method For The Model Unicellular Green Algae Chlamydomonas Reinhardtii, Truyen N. Quach, Shirley J. Sato, Mark R. Behrens, Paul N. Black, Concetta C. Dirusso, Heriberto D. Cerutti, Tom Elmo Clemente
Center for Plant Science Innovation: Faculty and Staff Publications
A reliable and simple Agrobacterium-mediated transformation system for the unicellular green algae model organism Chlamydomonas reinhardtii has been developed. The protocol has been successfully employed with both neomycin phosphotransferase II (nptII) and the phleomycin resistance (bleI) genes coupled with the selective agents paromomycin and zeocin, respectively. A set of binary vectors were assembled that carry the selectable marker cassettes under control either of the Rbcs2 alone or fused to the HSP270A leader sequence, PsaD, or ß-tubulin2 promoters. The corresponding T-DNA elements also harbored a cassette with a codon-optimized version of yellow fluorescence protein (YFP) under …
Evaluation Of The Escherichia Coli Threonine Deaminase Gene As A Selectable Marker For Plant Transformation, A. Ebmeier, L. Allison, Heriberto D. Cerutti, Thomas E. Clemente
Evaluation Of The Escherichia Coli Threonine Deaminase Gene As A Selectable Marker For Plant Transformation, A. Ebmeier, L. Allison, Heriberto D. Cerutti, Thomas E. Clemente
Center for Plant Science Innovation: Faculty and Staff Publications
The initial step in the synthesis of isoleucine (Ile) is the conversion of threonine to α-ketobutyrate. This reaction is carried out by threonine deaminase (TD), which is feedback-regulated by Ile. Mutations in TD that manifest insensitivity to Ile feedback inhibition result in intracellular accumulation of Ile. Previous reports have shown that in planta expression of the wild-type Escherichia coli TD, ilvA, or an Ile-insensitive mutant designated ilvA-466, increased cellular concentrations of Ile. A structural analog of Ile, L-O-methylthreonine (OMT), is able to compete effectively with Ile during translation and induce cell death. It has been postulated that OMT …