A yeast DNA recombination system facilitates the transference and expression of entire heterologous metabolic pathways into plant genomes.
Researchers have adapted a DNA recombination system from yeasts to facilitate the construction of large stretches of DNA and their introduction into plant genomes. This technology, called jStack, advances the engineering of new, complex functionality into plants by enabling the expression of heterologous multigene pathways.
Engineering more productive and resilient crops requires the introduction of new biological functions into plants. Often, multiple genes from different organisms need to be transferred to a given crop to provide new desirable properties, but assembling and introducing multiple genes into plant crops is difficult. The jStack technology will make it easier to combine genes from different sources and incorporate them into the genome of engineered crops to improve their performance.
Researchers at Lawrence Berkeley National Laboratory modified plasmid vectors commonly used for plant transformation so that they can be replicated and selected in yeasts, in addition to the Escherichia coli and plant hosts, to create a multigene plant transformation system called jStack. The system also includes yeast DNA sequences required for homologous recombination so that multiple DNA elements can be assembled into a single DNA molecule in yeast intermediary hosts in vivo. The resulting recombinant vectors can then be selected and purified for introduction in the desired plant host. In an attempt to standardize plant genetic engineering, the jStack system was designed to be compatible with commonly used cloning systems. Furthermore, a publicly available library of over 100 compatible promoters, genes, and terminator sequences was created to encourage collaboration and innovation within the plant synthetic biology community. The utility of the jStack technology was validated by introducing the entire pathway of the pigment violacein from the soil bacterium Chromobacterium violaceum into a model plant, as well as the metabolic pathway required to produce bisabolene, a precursor to bisabolane and a potential biodiesel component.
Contacts (BER PM)
DOE Office Biological and Environmental Research
Joint BioEnergy Institute
Biological Systems and Engineering Division
Lawrence Berkeley National Laboratory
This work was supported by the Office of Biological and Environmental Research within the U.S. Department of Energy’s Office of Science Early Career Research Program award to D. Loqué, and by contract DE-AC02-05CH11231.
Shih, P., K. Vuu, N. Mansoori, L. Ayad, K. Louie, B. Bowen, T. Northen, and D. Loqué. 2016. “A Robust Gene-Stacking Method Utilizing Yeast Assembly for Plant Synthetic Biology,” Nature Communications 7(13215), DOI: 10.1038/ncomms13215. (Reference link)
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