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Updated: Jun 28, 2026

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

Published on: February 9, 2015

Gateway cloning for protein expression.

Dominic Esposito¹, Leslie A Garvey, Chacko S Chakiath

¹Protein Expression Laboratory, National Cancer Institute, SAIC-Frederick, Frederick, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)

|November 7, 2008

Summary

This summary is machine-generated.

Recombinational cloning, especially the Gateway system, accelerates protein expression clone generation. This overcomes bottlenecks in producing soluble, high-yield proteins, making research more efficient and cost-effective.

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Published on: September 22, 2011

Area of Science:

Molecular Biology
Protein Expression
Biotechnology

Background:

Protein production is often limited by the inefficient generation of expression clones.
Producing functional human proteins in heterologous systems is challenging due to poor expression, aggregation, or purification issues.
Traditional cloning methods are time-consuming, costly, and often require multiple steps and redesigns.

Purpose of the Study:

To highlight the challenges in generating functional protein expression clones.
To introduce recombinational cloning as a solution to these challenges.
To emphasize the flexibility and efficiency of the Gateway system for protein expression.

Main Methods:

Discusses the limitations of traditional gene cloning for protein expression.
Introduces recombinational cloning techniques.
Highlights the Gateway recombinational cloning system.

Main Results:

Recombinational cloning enables rapid, parallel generation of expression clones.
The Gateway system offers high flexibility for protein expression.
This approach reduces costs and time associated with protein production pipelines.

Conclusions:

Recombinational cloning significantly improves protein expression workflows.
The Gateway system provides a robust and efficient method for generating expression clones.
This technology facilitates both low- and high-throughput protein production.