Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, MI, USA.
Current address:, MelaTech, LLC, Baltimore, Maryland, United States.
Methods Mol Biol. 2024;2760:21-34. doi: 10.1007/978-1-0716-3658-9_2.
As the field of plant synthetic biology continues to grow, Agrobacterium-mediated transient expression has become an essential method to rapidly test pathway candidate genes in a combinatorial fashion. This is especially important when elucidating and engineering more complex pathways to produce commercially relevant chemicals like many terpenoids, a widely diverse class of natural products of often industrial relevance. Agrobacterium-mediated transient expression has facilitated multiplex expression of recombinant and modified enzymes, including synthetic biology approaches to compartmentalize the biosynthesis of terpenoids subcellularly. Here, we describe methods on how to deploy Agrobacterium-mediated transient expression in Nicotiana benthamiana to rapidly develop terpenoid pathways and compartmentalize terpenoid biosynthesis within plastids, the cytosol, or at the surface of lipid droplets.
随着植物合成生物学领域的不断发展，农杆菌介导的瞬时表达已成为一种重要的方法，可以快速以组合方式测试途径候选基因。当阐明和工程更复杂的途径以生产具有商业相关性的化学物质时，如许多萜类化合物，这一点尤其重要，萜类化合物是一类广泛多样的天然产物，通常具有工业相关性。农杆菌介导的瞬时表达促进了重组和修饰酶的多重表达，包括将萜类化合物生物合成分区化到亚细胞水平的合成生物学方法。在这里，我们描述了如何在本氏烟中利用农杆菌介导的瞬时表达来快速开发萜类化合物途径，并将萜类化合物生物合成分区化到质体、细胞质或脂滴表面。
