Site-directed mutagenesis and saturation mutagenesis for the functional study of transcription factors involved in plant secondary metabolite biosynthesis

Sitakanta Pattanaik, Joshua R. Werkman, Que Kong, Ling Yuan

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

16 Scopus citations

Abstract

Regulation of gene expression is largely coordinated by a complex network of interactions between transcription factors (TFs), co-factors, and their cognate cis-regulatory elements in the genome. TFs are multidomain proteins that arise evolutionarily through protein domain shuffling. The modular nature of TFs has led to the idea that specific modules of TFs can be re-designed to regulate desired gene(s) through protein engineering. Utilization of designer TFs for the control of metabolic pathways has emerged as an effective approach for metabolic engineering. We are interested in engineering the basic helix-loop-helix (bHLH, Myc-type) transcription factors. Using site-directed and saturation mutagenesis, in combination with efficient and high-throughput screening systems, we have identified and characterized several amino acid residues critical for higher transactivation activity of a Myc-like bHLH transcription factor involved in anthocyanin biosynthetic pathway in plants. Site-directed and saturation mutagenesis should be generally applicable to engineering of all TFs.

Original languageEnglish
Title of host publicationPlant Secondary Metabolism Engineering
Subtitle of host publicationMethods and Applications
EditorsArthur Germano Fett-Neto
Pages47-57
Number of pages11
DOIs
StatePublished - 2010

Publication series

NameMethods in Molecular Biology
Volume643
ISSN (Print)1064-3745

Keywords

  • saturation mutagenesis
  • site-directed mutagenesis
  • Transcription factors
  • transient protoplast expression
  • yeast one-hybrid

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint

Dive into the research topics of 'Site-directed mutagenesis and saturation mutagenesis for the functional study of transcription factors involved in plant secondary metabolite biosynthesis'. Together they form a unique fingerprint.

Cite this