[Fusion expression of human thymosin alpha 1 in Escherichia coli].
Xiu. Zhao-Yang ZY; Yu. Ying Y; Chen. Chang-Qing CQ
Key Findings
- A GST‑Thymosin‑alpha‑1 fusion protein is expressed in E. coli at 35‑40% of total cellular protein
- Cleavage of the fusion yields roughly 0.2 g/L of pure thymosin‑alpha‑1
- Mass‑spec and activity tests confirm the recombinant peptide is biologically identical to the native form
Practical Outcomes
- The protocol offers a relatively high‑yield, low‑cost way for DIY biohackers to make thymosin‑alpha‑1 in‑house, provided they have access to basic molecular‑biology tools (cloning, fermentation, purification). It doesn’t give dosing or safety data, so users must still rely on existing human studies for how to use the peptide.
Summary
Scientists built a bacteria strain that can make the peptide thymosin‑alpha‑1, then cut it out to get about 0.2 g of pure, active peptide per litre of culture. The process is fairly efficient and the product works like the natural version, meaning hobby labs could potentially produce their own supply if they have the right equipment.
Abstract
Engineering E. coli strain, BL21 (DE3)/pGEX-4T-human Thymosin alpha 1, was constructed by oligonucleotide annealing and PCR amplifying the target gene, then ligating it with pGEX-4T-3 vector and transferring into BL21 host. The yield of fusion protein of GST-Thymosin alpha 1 expressed from BL21 (DE3)/pGEX-4T-thymosin alpha 1 is about 35%-40% of total protein after fermentation. Following the simple cut of thrombin or CNBr, about 0.2 g/L thymosin alpha 1 can be harvested. The product is checked by MS and activity test, which indicates that the recombinant product has full biological activity of native thymosin alpha 1.
Study Information
pubmed
2002