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Related Concept Videos

Genome Annotation and Assembly03:36

Genome Annotation and Assembly

The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.

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Related Experiment Video

Updated: Jun 21, 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

Protocol for connector-enabled multigene pathway assembly using Golden Gate Assembly and yeast recombination.

Min-Jun Seong1, Jonghyeok Shin2, Dae-Hee Lee3

  • 1Synthetic Biology Research Center and Korea Biofoundry, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 34141, Republic of Korea; Department of Biosystems and Bioengineering, KRIBB School of Biotechnology, University of Science and Technology (UST), Daejeon 34113, Republic of Korea.

STAR Protocols
|June 19, 2026
PubMed
Summary

This study introduces a new method for building multiple gene pathways in yeast. It simplifies complex genetic engineering, making it faster and easier for researchers to create new yeast strains.

Keywords:
Biotechnology and bioengineeringMolecular BiologySystems biology

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Yeast Genetics

Background:

  • Multigene pathway construction is crucial for metabolic engineering and synthetic biology.
  • Traditional methods can be complex, time-consuming, and require specialized expertise.
  • Standardized and efficient protocols are needed to accelerate yeast-based research.

Purpose of the Study:

  • To present a streamlined protocol for multigene pathway construction in Saccharomyces cerevisiae.
  • To integrate Golden Gate Assembly with yeast homologous recombination for enhanced efficiency.
  • To reduce complexity and operator dependence in assembling multiple transcriptional units.

Main Methods:

  • Developed a connector-enabled workflow for assembling Level 1 transcriptional unit (TU) fragments.
  • Utilized standardized connectors for seamless Golden Gate Assembly.
  • Co-transformed assembled TUs into Saccharomyces cerevisiae for Level 2 multigene plasmid formation.

Main Results:

  • The protocol significantly reduces procedural complexity compared to conventional methods.
  • Minimized the dependence on specialized operator expertise.
  • Substantially accelerated workflow timelines for multigene construct assembly.

Conclusions:

  • The presented protocol offers a simplified, faster, and more accessible approach for multigene pathway construction in yeast.
  • This method facilitates rapid assembly of complex genetic constructs in Saccharomyces cerevisiae.
  • The connector-enabled workflow enhances the efficiency and reproducibility of yeast synthetic biology applications.