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Modifying Baculovirus Expression Vectors to Produce Secreted Plant Proteins in Insect Cells

Modifying Baculovirus Expression Vectors to Produce Secreted Plant Proteins in Insect Cells

Published on: August 20, 2018

GPCR expression using baculovirus-infected Sf9 cells.

Amanda L Aloia¹, Richard V Glatz, Edward J McMurchie

¹School of Biology, Flinders University, Bedford Park, SA, Australia.

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

|June 11, 2009

Summary

Recombinant baculoviruses enable high-yield expression of functional G protein-coupled receptors (GPCRs) in insect cells. This method allows for protein detection, quantification, and modulation of expression levels via ligand addition.

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Transient Expression of Foreign Genes in Insect Cells (sf9) for Protein Functional Assay

Transient Expression of Foreign Genes in Insect Cells (sf9) for Protein Functional Assay

Published on: February 22, 2018

Area of Science:

Molecular Biology
Biotechnology
Pharmacology

Background:

Insect cell expression systems using recombinant baculoviruses are widely adopted for G protein-coupled receptor (GPCR) research.
This system offers high yields of functional receptors, capacity for post-translational modifications, and scalability for large-scale cultures.

Purpose of the Study:

To present generic methods for expressing GPCRs in insect cells via baculovirus infection.
To demonstrate the expression and characterization of a specific mammalian M(2) muscarinic receptor in Sf9 cells.

Main Methods:

Utilizing recombinant baculoviruses for GPCR expression in insect cell cultures.
Maintaining insect cell cultures and employing polyhedrin promoter-driven expression.
Detecting and quantifying expressed receptors using radiolabeled ligand binding assays.

Main Results:

Maximal receptor expression was observed approximately 72 hours post-infection.
Expressed M(2) muscarinic receptor was successfully detected and quantified.
Ligand addition to infected insect cell cultures altered receptor expression levels.

Conclusions:

The baculovirus expression system provides a robust platform for producing functional GPCRs in insect cells.
This method facilitates receptor detection, quantification, and manipulation for further study.