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Updated: May 26, 2026

Functional Cloning Using a Xenopus Oocyte Expression System

Functional Cloning Using a Xenopus Oocyte Expression System

Published on: January 30, 2016

Efficient selection of transgenic mouse embryos using EGFP as a marker gene.

M Kato¹, K Yamanouchi, M Ikawa

¹Laboratory of Applied Genetics, Graduate School of Agricultural and Life Sciences, University of Tokyo, Japan.

Molecular Reproduction and Development

|July 28, 1999

Summary

This summary is machine-generated.

Enhanced Green Fluorescent Protein (EGFP) reliably identifies transgenic embryos. Uniform EGFP expression in all cells is crucial for confirming successful gene transfer and obtaining transgenic animals.

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Published on: August 24, 2022

Area of Science:

Molecular Biology
Developmental Biology
Genetics

Background:

Efficiently identifying transgenic embryos is critical for genetic research and animal model development.
Traditional methods for transgenic identification can be time-consuming and may impact embryo viability.

Purpose of the Study:

To establish and validate a reliable method for selecting transgenic preimplantation embryos using Enhanced Green Fluorescent Protein (EGFP) as a marker.
To assess the impact of EGFP expression monitoring on embryo developmental potential.

Main Methods:

Pronuclear microinjection of the CMV/beta-actin/EGFP fusion gene into mouse embryos.
In vitro culture of embryos to morula or blastocyst stages.
Fluorescence microscopy for EGFP expression observation.
Modified PCR analysis (Dpn I and Bal 31 digestion) for transgenic confirmation.

Main Results:

EGFP expression was readily detectable via fluorescence microscopy.
No significant damage to embryo developmental ability was observed after 30 min of EGFP excitation.
Embryos with uniform EGFP expression were confirmed as transgenic, while mosaic expression often indicated non-transgenic status.
Transgenic offspring rates were 77% for uniformly EGFP-expressing embryos versus 21.4% for mosaic ones.

Conclusions:

EGFP is a highly useful and reliable marker for selecting transgenic embryos.
Confirming uniform EGFP expression across all cells is essential for accurate identification of truly transgenic animals.