Biosynthesis of Proteins Inside Liposomes
Protein expression is the most complex metabolic reaction that has been encapsulated in liposomes, mainly as an intermediate step toward the synthesis of minimal semisynthetic cells. Although there are different experimental approaches to achieving the synthesis of proteins inside liposomes and it is therefore not possible to give a standard recipe, all methods follow a general strategy, which is briefly discussed. On this basis, we provide general indications for designing and realizing protein-expressing liposomes. Our approach for the green fluorescent protein expression inside 200-nm extruded vesicles is described in detail.
Key wordsCell-free protein expression PURESYSTEM Enhanced green fluorescent protein (EGFP) Minimal cell Synthetic biology
This work was carried out within the SYNTHCELLS project (Approaches to the Bioengineering of Synthetic Minimal Cells, EU Grant #FP6-043359) and further supported by the Human Frontiers Science Program.
- 4.Luisi PL (2006) The emergence of life: from chemical origin to synthetic biology. Cambridge University Press, CambridgeGoogle Scholar
- 14.Souza T, Stano P, Luisi PL (2009) The minimal size of liposome-based model cells brings about a remarkably enhanced entrapment and protein synthesis. ChemBioChem 10: 1056-1063Google Scholar
- 15.Kuruma Y, Stano P, Ueda T, Luisi PL (2009) A synthetic biology approach to the construction of membrane proteins in semi-synthetic minimal cells. Biochim. Biophys. Acta 1788: 567-574Google Scholar
- 28.Cevc G (ed) (1993) Phospholipids handbook. Marcel Dekker Inc, New YorkGoogle Scholar
- 31.PROMEGA Product # L5001, technical NOTES, # TB285Google Scholar
- 32.Harris DH (2003) Quantitative chemical analysis. W. H. Freeman and Co., New YorkGoogle Scholar