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Related Concept Videos

Reporter Genes02:11

Reporter Genes

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Updated: Apr 1, 2026

Optimization and Utilization of Agrobacterium-mediated Transient Protein Production in Nicotiana
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Comprehensive Optimization of Agroinfiltration in Transient Expression System in Nicotiana tabacum with RUBY Visible

M R Sorokina1, Yu A Yugay1, A I Degtyarenko1

  • 1Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch, Russian Academy of Sciences, Vladivostok, Russia.

Doklady Biological Sciences : Proceedings of the Academy of Sciences of the USSR, Biological Sciences Sections
|March 30, 2026
PubMed
Summary

Optimizing Agrobacterium-mediated transient expression in Nicotiana tabacum enhances gene expression. This protocol improves efficiency for plant biotechnology applications like functional genomics and metabolic engineering.

Keywords:
Nicotiana tabacumRUBYagrobacterium-mediated transient expressionbetalain

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Area of Science:

  • Plant Biotechnology
  • Functional Genomics
  • Molecular Biology

Background:

  • Agrobacterium-mediated transient expression is crucial for plant biotechnology and functional genomics.
  • Expression efficiency and reproducibility are influenced by various factors, necessitating optimization.

Purpose of the Study:

  • To optimize agroinfiltration conditions for enhanced betalain gene expression in Nicotiana tabacum.
  • To improve the efficiency and reproducibility of transient gene expression.

Main Methods:

  • Evaluated effects of bacterial suspension concentration, infiltration buffer composition, and delivery methods.
  • Utilized the RUBY visual reporter system for assessing gene expression.
  • Tested various buffer components including acetosyringone, MES, sucrose, and magnesium chloride.

Main Results:

  • Optimal conditions included OD600 = 1.0 bacterial suspension.
  • The best buffer contained 50 μM acetosyringone, 10 mM MES, 0.5% sucrose, and 10 mM MgCl2.
  • Preliminary leaf micropuncture followed by vacuum infiltration yielded the highest expression.

Conclusions:

  • The developed protocol significantly enhances transient expression efficiency in Nicotiana tabacum.
  • This optimized method is applicable to gene functional analysis, metabolic engineering, and biopharmaceutical production in plants.