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Related Experiment Video

Updated: Jun 22, 2025

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

TOPO Cloning for Efficient Plasmid Construction.

Maya Yovcheva¹, Kenneth W Thompson²

¹Thermo Fisher Scientific, Frederick, MD, USA. Maya.Yovcheva@thermofisher.com.

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

|July 1, 2024

Summary

This summary is machine-generated.

TOPO cloning offers a fast way to create recombinant plasmids for the Bac-to-Bac Baculovirus Expression System. This method simplifies gene cloning for protein expression in insect cells.

Keywords:

Baculovirus expression vector system Gene of interest TOPO cloning

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Last Updated: Jun 22, 2025

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Published on: January 8, 2015

Area of Science:

Molecular Biology
Recombinant DNA Technology
Protein Expression Systems

Background:

The Bac-to-Bac system is a powerful tool for high-level protein expression in insect cells.
Efficient generation of recombinant baculovirus is crucial for successful protein production.
Traditional cloning methods can be time-consuming and labor-intensive.

Purpose of the Study:

To describe a streamlined method for generating recombinant plasmids for the Bac-to-Bac system.
To highlight the advantages of using TOPO cloning in this context.

Main Methods:

Utilizes TOPO cloning technology for rapid insertion of genes of interest into a transfer vector.
The resulting recombinant plasmid is then used for transformation into competent E. coli.
Transformed E. coli are selected and cultured to obtain the desired plasmid DNA.

Main Results:

Demonstrates the efficiency and speed of TOPO cloning for generating the required recombinant plasmid.
Confirms successful generation of plasmids suitable for subsequent steps in the Bac-to-Bac system.

Conclusions:

TOPO cloning provides a highly efficient and user-friendly approach for preparing plasmids for the Bac-to-Bac Baculovirus Expression System.
This method significantly accelerates the workflow for recombinant protein expression studies.