Abstract
Production of a protein of interest in bacteria and its purification from bacterial lysates are valuable tools for the purification of larger amounts of recombinant proteins. The low cost of culturing, and the rapid cell growth of bacteria make this host a good choice for protein production, but the folding and function of the purified protein might be altered due to the production of a eukaryotic protein in a prokaryotic host. Here, we provide a purification method for the purification of gamma (γ)-tubulin (TUBG) from soluble fractions of Escherichia (E.) coli lysates using affinity tags.
This protocol describes a method that purifies soluble GST-TUBG1 from bacteria.
Of the three tested induction conditions, the highest yield of recombinant GST-TUBG1 was obtained after the induction of E. coli with isopropyl-D-1-thiogalactopyranoside (IPTG) for 1 h at 37°C followed by overnight incubation at room temperature.
In comparison with other methodologies [1], the technique described here retrieves larger amounts of recombinant γ-tubulin from small-scale expression cultures.
This protocol describes a method that purifies soluble GST-TUBG1 from bacteria.
Of the three tested induction conditions, the highest yield of recombinant GST-TUBG1 was obtained after the induction of E. coli with isopropyl-D-1-thiogalactopyranoside (IPTG) for 1 h at 37°C followed by overnight incubation at room temperature.
In comparison with other methodologies [1], the technique described here retrieves larger amounts of recombinant γ-tubulin from small-scale expression cultures.
| Original language | English |
|---|---|
| Article number | 101517 |
| Pages (from-to) | 1-8 |
| Journal | MethodsX |
| Volume | 8 |
| DOIs | |
| Publication status | Published - 2021 |
Subject classification (UKÄ)
- Biochemistry and Molecular Biology