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Related Experiment Videos

Transcriptional interference in transgenic plants.

I Ingelbrecht1, P Breyne, K Vancompernolle

  • 1Laboratorium voor Genetica, Universiteit Gent, Belgium.

Gene
|December 30, 1991
PubMed
Summary

Agrobacterium-mediated plant transformation preferentially integrates T-DNA into transcribed regions. Transcriptional readthrough significantly reduces downstream gene expression, but poly(A) signals can restore it.

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Area of Science:

  • Plant molecular biology
  • Gene expression regulation
  • Agrobacterium-mediated transformation

Background:

  • Agrobacterium tumefaciens mediated transformation results in T-DNA integration into plant genomes.
  • Promoterless marker genes integrated into plant genomes show a high frequency of gene fusion, suggesting preferential integration into transcribed regions.

Purpose of the Study:

  • To investigate the role of transcriptional activity in gene expression variation among independent transgenic plants.
  • To determine the impact of transcriptional readthrough on downstream gene expression.
  • To assess the role of poly(A) signals in mitigating readthrough effects.

Main Methods:

  • Utilized hybrid gene constructions to study gene expression.
  • Employed transient expression assays and analysis of transgenic plants.

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  • Conducted enzymatic analysis of calli to quantify gene expression variation.

    • Main Results:

    • Transcriptional readthrough into oppositely oriented downstream genes substantially reduced their expression.
    • Polyadenylation (poly(A)) signals and terminators effectively blocked readthrough and restored gene expression.
    • Separating genes with promoter and terminator elements reduced expression variation in transgenic plants.

    Conclusions:

    • Transcriptional activity at the integration site contributes to expression variability in transgenic plants.
    • Proper gene termination elements are crucial for stable and predictable gene expression in transgenic systems.
    • Understanding T-DNA integration and gene expression dynamics is vital for successful plant genetic engineering.