Precise and reliable control of gene expression in Agrobacterium tumefaciens

Yuanchao Qian, Wentao Kong, Ting Lu

Research output: Contribution to journalArticlepeer-review

Abstract

Agrobacterium tumefaciens is a soil-borne bacterium that is known for its DNA delivery ability and widely exploited for plant transformation. Despite continued interest in improving the utility of the organism, the lack of well-characterized engineering tools limits the realization of its full potential. Here, we present a synthetic biology toolkit that enables precise and effective control of gene expression in A. tumefaciens. We constructed and characterized six inducible expression systems. Then, we optimized the one regulated by cumic acid through amplifier introduction and promoter engineering and evaluated its 15 cognate promoters. To establish fine-tunability, we constructed a series of spacers and a promoter library to systematically modulate both translational and transcriptional rates. We finally demonstrated the application of the tools by co-expressing genes with altered expression levels using a single signal. This study provides precise expression tools for A. tumefaciens, facilitating rational engineering of the bacterium for advanced plant biotechnological applications.

Original languageEnglish (US)
Pages (from-to)3962-3972
Number of pages11
JournalBiotechnology and bioengineering
Volume118
Issue number10
DOIs
StatePublished - Oct 2021

Keywords

  • Agrobacterium tumefaciens
  • gene expression
  • promoter library
  • synthetic biology
  • VirG mutants

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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