Overexpression of soluble human thymosin alpha 1 in Escherichia coli.
Chen. Pei-Fu PF; Zhang. Hong-Ying HY; Fu. Geng-Feng GF; Xu. Gen-Xing GX; Hou. Ya-Yi YY
Key Findings
- BL21/pET‑28a system gave the highest soluble expression, up to 70% of total bacterial proteins
- Purified fusion protein represented 2.53% of the wet bacterial pellet weight
- Purity of the final product reached 94.5% by SDS‑PAGE
Practical Outcomes
- The method could be used by labs or advanced DIY biohackers to produce thymosin‑alpha‑1 more cheaply and at scale. However, it requires cloning, bacterial culture, and chromatography equipment, so it isn’t a ready‑to‑use protocol for most users.
Summary
Scientists figured out a way to make a lot of the peptide thymosin‑alpha‑1 in common lab bacteria (E. coli) using a specific DNA construct, getting it mostly in a soluble form and purifying it to high purity. This shows a practical recipe for producing the peptide in bulk, though it still needs standard molecular‑biology tools.
Abstract
Synthesized gene of human thymosin alpha 1 (Talpha1) was inserted into pET-28a, pET-9c, pThioHis B, pGEX-2T or pBV222 and then inductively expressed in strains of Escherichia coli. Among the five expression systems, the BL21/pET-28a system provides the highest expression level of fusion protein in a soluble form, which is up to 70% of total expressed bacterial proteins as visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The resulting fusion protein purified through nickel affinity chromatography accounts for 2.53% of the wet bacterial pellet weight and reaches 94.5% purity by SDS-PAGE. These results indicate the potential of this expression system for high-throughput production of recombinant Talpha1.
Study Information
pubmed
2005