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Related Experiment Video

Updated: Jun 1, 2026

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
08:31

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Published on: February 5, 2021

Simple cloning strategy using GFPuv gene as positive/negative indicator.

Hiromi Miura1, Hidetoshi Inoko, Ituro Inoue

  • 1Department of Molecular Life Science, Division of Basic Medical Science and Molecular Medicine, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan.

Analytical Biochemistry
|May 24, 2011
PubMed
Summary

Researchers developed new cloning vectors for gene functional analysis. These vectors simplify clone detection and expression vector construction using a GFPuv gene indicator and type IIS restriction enzyme, aiding postgenomic studies.

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Area of Science:

  • Molecular Biology
  • Gene Expression Analysis

Background:

  • Gene functional analysis relies heavily on expression vector construction.
  • The postgenomic era demands simple and efficient cloning systems.

Purpose of the Study:

  • To develop novel cloning vectors for gene gain- or loss-of-function studies.
  • To create a simplified cloning procedure using a positive/negative indicator.

Main Methods:

  • Utilized the GFPuv gene as a reporter for successful cloning.
  • Employed a type IIS restriction enzyme for precise vector manipulation.
  • Integrated GFPuv gene and type IIS enzyme for a streamlined cloning process.

Main Results:

  • Successfully developed cloning vectors enabling easy detection of correct clones.

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Last Updated: Jun 1, 2026

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
08:31

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Published on: February 5, 2021

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  • Demonstrated a simplified procedure for obtaining functional expression vectors.
  • The combined use of GFPuv and type IIS restriction enzyme proved effective.

    • Conclusions:

    • The developed cloning vectors offer a simple and efficient method for gene functional analysis.
    • This system facilitates the construction of expression vectors, crucial for postgenomic research.
    • The GFPuv indicator and type IIS enzyme combination enhances cloning workflow.