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Related Experiment Video

Updated: Jun 1, 2026

An Efficient Method for Adenovirus Production
10:06

An Efficient Method for Adenovirus Production

Published on: June 10, 2021

Methods to construct recombinant adenovirus vectors.

Miguel Chillon1, Ramon Alemany

  • 1Biochemistry and Molecular Biology Department, Laboratory of Gene Therapy for Autoimmune Diseases, CBATEG, Universitat Autònoma Barcelona, Barcelona, Spain. Miguel.chillon@uab.es

Methods in Molecular Biology (Clifton, N.J.)
|May 19, 2011
PubMed
Summary

Homologous recombination in microorganisms offers an efficient method for adenovirus gene insertion. This chapter details popular Escherichia coli and yeast-based procedures for creating recombinant adenovirus, ensuring quality before large-scale production.

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Area of Science:

  • Molecular Biology
  • Virology
  • Biotechnology

Background:

  • Adenovirus vectors are crucial tools in gene therapy and vaccine development.
  • Efficiently introducing genes into the adenovirus genome is essential for creating effective vectors.
  • Homologous recombination (HR) in microorganisms provides a robust system for genetic manipulation of large viral genomes.

Purpose of the Study:

  • To describe and compare popular homologous recombination methods for generating recombinant adenoviruses.
  • To highlight key differences in plasmid requirements and selection systems for recombinant adenovirus identification.
  • To outline the critical quality control steps for adenovirus pre-stocks prior to large-scale amplification.

Main Methods:

  • Detailed protocols for two homologous recombination procedures in Escherichia coli.

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Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5
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Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5

Published on: January 28, 2016

Related Experiment Videos

Last Updated: Jun 1, 2026

An Efficient Method for Adenovirus Production
10:06

An Efficient Method for Adenovirus Production

Published on: June 10, 2021

Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5
13:17

Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5

Published on: January 28, 2016

  • Description of one homologous recombination procedure in yeast.
  • Methods for analyzing adenovirus genome identity and integrity.
  • Generation and quality assessment of viral pre-stocks in permissive human cells.

    • Main Results:

    • Successful generation of recombinant adenoviruses using described HR methods.
    • Identification of key variations in plasmid design and selection strategies impacting efficiency.
    • Establishment of protocols for confirming viral genome integrity and excluding replication-competent particles.

    Conclusions:

    • Homologous recombination in microorganisms is a highly efficient system for adenovirus genome engineering.
    • The choice of HR procedure impacts plasmid needs and recombinant identification speed.
    • Rigorous analysis of viral pre-stocks is critical for ensuring safety and potency in large-scale adenovirus production.