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Related Experiment Videos

A recombination-efficient baculovirus vector for simultaneous expression of multiple genes

U Chatterji1, R Ahmad, B Venkaiah

  • 1Eukaryotic Gene Expression Laboratory, National Institute of Immunology, New Delhi, India.

Gene
|June 1, 1996
PubMed
Summary

A novel baculovirus expression vector, pBacUCmP3, enables high-frequency cloning and expression of multiple foreign genes in eukaryotic cells. This tool facilitates simultaneous protein synthesis at different time points for diverse applications.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Gene Expression Systems

Background:

  • The baculovirus expression system is a powerful tool for heterologous gene expression in eukaryotic environments.
  • Efficient cloning and expression of multiple genes are crucial for various biotechnological applications.

Purpose of the Study:

  • To construct and characterize a multiple expression transfer vector (pBacUCmP3) for simultaneous expression of three independent foreign genes.
  • To demonstrate the utility of the vector for expressing reporter genes under different promoter controls.

Main Methods:

  • Construction of the pBacUCmP3 vector containing dual promoters (Autographa californica nuclear polyhedrosis virus very late gene promoter and Drosophila melanogaster heat-shock protein 70 promoter).
  • Co-transfection of the vector with linearized viral DNA.

Related Experiment Videos

  • Expression analysis of reporter genes (beta-subunit of human chorionic gonadotropin hormone, firefly luciferase, bacterial beta-galactosidase) post-infection.

    • Main Results:

    • High frequencies of recombinant progeny viruses were obtained.
    • Beta-galactosidase expression was controlled by the hsp70 promoter.
    • Beta hCG and luciferase expression followed the polyhedrin promoter activation profile.
    • Differential gene expression timing was confirmed.

    Conclusions:

    • The pBacUCmP3 vector facilitates high-frequency cloning and expression of multiple genes.
    • The vector allows for temporal control of protein synthesis using distinct promoters.
    • This tool is valuable for applications requiring the synthesis of multiple proteins at different time points.