Synthesis of Preparative Amounts of Biologically Active Interleukin-6 Using a Continuous-Flow Cell-Free Translation System
✍ Scribed by E.V. Volyanik; A. Dalley; I.A. Mckay; I. Leigh; N.S. Williams; S.A. Bustin
- Publisher
- Elsevier Science
- Year
- 1993
- Tongue
- English
- Weight
- 457 KB
- Volume
- 214
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Cell-free synthesis of preparative amounts of interleukin-6 (IL-6) in a wheat germ extract is described. Following transcription of uncapped mRNA by SP6 polymerase, protein was synthesized in a continuous flow of translation components. The system remained active for at least (50 \mathrm{~h}), producing (1.25 \times 10^{8}) units of IL-6. Recovered IL-6 was (>80 %) pure, reacted with anti-IL-6 polyclonal antiserum in Western blots, and had a specific activity comparable to recombinant IL-6 purified from Escherichia coli. This is the first time biologically active IL-6 has been prepared in preparative amounts using a continuous-flow cell-free translation system and confirms the feasibility of using enhanced in vitro translation for rapid synthesis of proteins from cloned templates. 01993 Academic Press, Inc.