Assembly of plant virus agroinfectious clones using biological material or DNA synthesis

STAR Protoc. 2022 Dec 16;3(4):101716. doi: 10.1016/j.xpro.2022.101716. Epub 2022 Sep 22.

Abstract

Infectious clone technology is universally applied for biological characterization and engineering of viruses. This protocol describes procedures that implement synthetic biology advances for streamlined assembly of virus infectious clones. Here, I detail homology-based cloning using biological material, as well as SynViP assembly using type IIS restriction enzymes and chemically synthesized DNA fragments. The assembled virus clones are based on compact T-DNA binary vectors of the pLX series and are delivered to host plants by Agrobacterium-mediated inoculation. For complete details on the use and execution of this protocol, please refer to Pasin et al. (2017, 2018) and Pasin (2021).

Keywords: Biotechnology and bioengineering; Genomics; Microbiology; Molecular biology; Plant sciences; Sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Plant Viruses* / genetics
  • Plants