Combining a Simple Method for DNA/RNA/Protein Co-Purification and Arabidopsis Protoplast Assay to Facilitate Viroid Research

Viruses. 2019 Apr 3;11(4):324. doi: 10.3390/v11040324.

Abstract

Plant-viroid interactions represent a valuable model for delineating structure-function relationships of noncoding RNAs. For various functional studies, it is desirable to minimize sample variations by using DNA, RNA, and proteins co-purified from the same samples. Currently, most of the co-purification protocols rely on TRI Reagent (Trizol as a common representative) and require protein precipitation and dissolving steps, which render difficulties in experimental handling and high-throughput analyses. Here, we established a simple and robust method to minimize the precipitation steps and yield ready-to-use RNA and protein in solutions. This method can be applied to samples in small quantities, such as protoplasts. Given the ease and the robustness of this new method, it will have broad applications in virology and other disciplines in molecular biology.

Keywords: DNA/RNA/protein co-purification; protoplast; viroid.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Arabidopsis / cytology*
  • Arabidopsis / genetics
  • Arabidopsis / virology
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / isolation & purification
  • Arabidopsis Proteins / metabolism
  • DNA, Plant / genetics
  • DNA, Plant / isolation & purification
  • DNA, Plant / metabolism
  • Genome, Plant
  • Host-Pathogen Interactions
  • Molecular Biology / methods*
  • Protoplasts / metabolism
  • Protoplasts / virology*
  • RNA, Plant / isolation & purification
  • RNA, Plant / metabolism
  • RNA, Viral / isolation & purification
  • RNA, Viral / metabolism
  • Viroids / genetics
  • Viroids / physiology*
  • Virus Replication

Substances

  • Arabidopsis Proteins
  • DNA, Plant
  • RNA, Plant
  • RNA, Viral