Related Experiment Video
Updated: Mar 8, 2026
08:57
Production of Recombinant PRMT Proteins using the Baculovirus Expression Vector System
Published on: July 17, 2021
8.4K
Expression and Purification of SH2 Domains Using Baculovirus Expression System
Mari Ogiue-Ikeda1, Kazuya Machida2
1Raymond and Beverly Sackler Laboratory of Genetics and Molecular Medicine, Department of Genetics and Genome Sciences, University of Connecticut School of Medicine, 400 Farmington Avenue, Farmington, CT, 06032, USA.
Methods in Molecular Biology (Clifton, N.J.)
|January 17, 2017
Summary
Bacterial expression of Src homology 2 (SH2) domains can yield insoluble proteins. The baculovirus-insect cell system offers an alternative for producing soluble and active SH2 domains.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Expression
Background:
- Recombinant proteins produced in bacterial systems often exhibit poor solubility, aggregation, and incorrect folding.
- This is a common challenge in protein research and biotechnology.
Purpose of the Study:
- To present a reliable protocol for expressing and purifying Src homology 2 (SH2) domains.
- To demonstrate the utility of baculovirus-insect cell expression systems for overcoming bacterial expression limitations.
Main Methods:
- Utilized the Bac-to-Bac baculovirus expression system for protein production.
- Employed cloning and purification strategies for GST-tagged SH2 domains.
Main Results:
- Successfully produced soluble and functionally active GST-tagged SH2 domains.
- Overcame the insolubility and aggregation issues typically encountered with bacterial expression.
Conclusions:
- The baculovirus-insect cell expression system is an effective alternative for producing soluble and active SH2 domains.
- This protocol facilitates the study of SH2 domain function and interactions.
