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Related Experiment Video

Updated: May 24, 2026

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
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Exploiting Multisite Gateway and pENFRUIT plasmid collection for fruit genetic engineering.

Leandro H Estornell1, Antonio Granell, Diego Orzaez

  • 1Instituto de Biología Molecular y Celular de Plantas, Valencia, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|February 22, 2012
PubMed
Summary

pENFRUIT utilizes MultiSite Gateway cloning for efficient genetic engineering in fruits. This method enables rapid assembly of gene expression and silencing cassettes for transgenic fruit research.

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Area of Science:

  • Plant Molecular Biology
  • Genetic Engineering
  • Biotechnology

Background:

  • MultiSite Gateway cloning enables combinatorial assembly of genetic elements like promoters, CDS, and terminators into gene expression or silencing cassettes.
  • Genetic engineering in fruits requires efficient methods for assembling and testing gene constructs.

Purpose of the Study:

  • To introduce pENFRUIT, a collection of MultiSite Triple Gateway Entry vectors designed for fruit genetic engineering.
  • To facilitate the combinatorial assembly of fruit-specific promoters and C-terminal tags with genes of interest.

Main Methods:

  • Utilized MultiSite Triple Gateway cloning methodology for vector assembly.
  • Developed a set of fruit-operating promoters and C-terminal tags compatible with the Gateway system.

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Last Updated: May 24, 2026

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  • Assembled gene constructs for ectopic expression or silencing in fruits.

    • Main Results:

    • pENFRUIT vectors enable efficient Gateway assembly of genetic elements for fruit engineering.
    • The system allows for the creation of diverse gene expression and silencing cassettes.
    • Constructs can be tested using stable transgenic plants or transient expression assays.

    Conclusions:

    • pENFRUIT provides a valuable toolkit for advancing genetic engineering in fruit crops.
    • The MultiSite Gateway cloning approach accelerates the testing of gene function and regulation in fruits.
    • This methodology supports the rapid development of improved fruit varieties.