ARGONAUTE Genes in Salvia miltiorrhiza: Identification, Characterization, and Genetic Transformation

Methods Mol Biol. 2017:1640:173-189. doi: 10.1007/978-1-4939-7165-7_12.

Abstract

Small RNA-mediated gene silencing is a vital regulatory mechanism in eukaryotes that requires ARGONAUTE (AGO) proteins. Salvia miltiorrhiza is a well-known traditional Chinese medicinal plant. Therefore, it is important to characterize S. miltiorrhiza AGO family genes as they may be involved in multiple metabolic pathways. This chapter introduces the detailed protocol for SmAGO gene prediction and molecular cloning. In addition, an Agrobacterium-mediated genetic transformation method for S. miltiorrhiza is presented. These methodologies can be used to functionally study SmAGO genes as well as other genes of interest in S. miltiorrhiza.

Keywords: ARGONAUTE; Gene identification; Genetic transformation; Salvia miltiorrhiza; Small RNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium / genetics
  • Argonaute Proteins / genetics*
  • Cloning, Molecular / methods*
  • Gene Expression Regulation, Plant
  • Genes, Plant
  • Plant Proteins / genetics*
  • Polymerase Chain Reaction / methods
  • Salvia miltiorrhiza / genetics*
  • Salvia miltiorrhiza / growth & development
  • Transformation, Genetic*

Substances

  • Argonaute Proteins
  • Plant Proteins