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Updated: Jun 3, 2026

The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions

The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions

Published on: December 15, 2012

Gene and genome construction in yeast.

Daniel G Gibson¹

¹The J. Craig Venter Institute, San Diego, California, USA.

Current Protocols in Molecular Biology

|April 8, 2011

Summary

Yeast can assemble large DNA fragments, including bacterial genomes, from single- or double-stranded DNA pieces in one step. This method enables the creation of complex DNA molecules for various research applications.

Area of Science:

Molecular Biology
Synthetic Biology
Genomics

Background:

The yeast Saccharomyces cerevisiae is a powerful model organism for genetic studies.
Efficient methods for DNA assembly are crucial for synthetic biology and genome engineering.

Purpose of the Study:

To present protocols for DNA assembly in yeast.
To demonstrate the capacity of yeast for assembling large and complex DNA molecules.

Main Methods:

Designing single-stranded oligonucleotides and double-stranded DNA fragments for assembly.
Transforming yeast (Saccharomyces cerevisiae) with designed DNA molecules.
Confirming the successful assembly of DNA molecules using various techniques.

Main Results:

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Last Updated: Jun 3, 2026

The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions

The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions

Published on: December 15, 2012

Genetic Engineering of an Unconventional Yeast for Renewable Biofuel and Biochemical Production

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Published on: September 20, 2016

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

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

Published on: February 5, 2021

Yeast can assemble multiple overlapping DNA molecules in a single transformation event.
Both single-stranded oligonucleotides and double-stranded DNA fragments can be utilized for assembly.
The system supports the assembly of gene-sized fragments up to complete bacterial genomes.

Conclusions:

Saccharomyces cerevisiae provides a versatile platform for large-scale DNA assembly.
The presented protocols facilitate the construction of complex DNA constructs, including entire genomes.
This technique has broad implications for synthetic biology, metabolic engineering, and genome research.