InteBac: An integrated bacterial and baculovirus expression vector suite

The successful production of recombinant protein for biochemical, biophysical, and structural biological studies critically depends on the correct expression organism. Currently, the most commonly used expression organisms for structural studies areEscherichia coli (similar to 70% of all PDB structures) and the baculovirus/ insect cell expression system (similar to 5% of all PDB structures). While insect cell expression is frequently successful for large eukaryotic proteins, it is relatively expensive and time-consuming compared toE. coliexpression. Frequently the decision to carry out a baculovirus project means restarting cloning from scratch. Here we describe an integrated system that allows simultaneous cloning intoE. coliand baculovirus expression vectors using the same PCR products. The system offers a flexible array of N- and C-terminal affinity, solubilization and utility tags, and the speed allows expression screening to be completed inE. coli, before carrying out time and cost-intensive experiments in baculovirus. Importantly, we describe a means of rapidly generating polycistronic bacterial constructs based on the hugely successful biGBac system, making InteBac of particular interest for researchers working on recombinant protein complexes.

Automated summary

The choice of expression organism is critical for successful production of recombinant proteins, with Escherichia coli and baculovirus/insect cell expression system being the most commonly used for structural studies. An integrated system allowing simultaneous cloning into both E. coli and baculovirus expression vectors can enhance experimental efficiency and flexibility.