BIOMAT Database Logo BIOMAT Database Logo

Synthesis of preparative amounts of biologically active interleukin-6 using a continuous-flow cell-free translation system. 1993

E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
Surgical Unit, London Hospital Medical College, University of London, Whitechapel, England.

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 h, producing 1.25 x 10(6) 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.

UI MeSH Term Description Entries
D007106 Immune Sera Serum that contains antibodies. It is obtained from an animal that has been immunized either by ANTIGEN injection or infection with microorganisms containing the antigen. Antisera,Immune Serums,Sera, Immune,Serums, Immune
D007700 Kinetics The rate dynamics in chemical or physical systems.
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D002455 Cell Division The fission of a CELL. It includes CYTOKINESIS, when the CYTOPLASM of a cell is divided, and CELL NUCLEUS DIVISION. M Phase,Cell Division Phase,Cell Divisions,Division Phase, Cell,Division, Cell,Divisions, Cell,M Phases,Phase, Cell Division,Phase, M,Phases, M
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D002470 Cell Survival The span of viability of a cell characterized by the capacity to perform certain functions such as metabolism, growth, reproduction, some form of responsiveness, and adaptability. Cell Viability,Cell Viabilities,Survival, Cell,Viabilities, Cell,Viability, Cell
D002474 Cell-Free System A fractionated cell extract that maintains a biological function. A subcellular fraction isolated by ultracentrifugation or other separation techniques must first be isolated so that a process can be studied free from all of the complex side reactions that occur in a cell. The cell-free system is therefore widely used in cell biology. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p166) Cellfree System,Cell Free System,Cell-Free Systems,Cellfree Systems,System, Cell-Free,System, Cellfree,Systems, Cell-Free,Systems, Cellfree
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D005813 Genes, Synthetic Biologically functional sequences of DNA chemically synthesized in vitro. Artificial Genes,Synthetic Genes,Artificial Gene,Gene, Artificial,Gene, Synthetic,Genes, Artificial,Synthetic Gene

Related Publications

E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
Preparative in vitro synthesis of bioactive human interleukin-2 in a continuous flow translation system.
October 1992, Biotechnology and applied biochemistry,
E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
Production of an enzymatic active protein using a continuous flow cell-free translation system.
September 1992, Journal of biotechnology,
E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
Preparative synthesis of globin in a continuous cell-free translation system from rabbit reticulocytes.
June 1989, Nucleic acids research,
E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
Requirement of continuous transcription for the synthesis of sufficient amounts of protein by a cell-free rapid translation system.
March 2002, Protein expression and purification,
E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
A continuous cell-free protein synthesis system for coupled transcription-translation.
August 1991, Journal of biochemistry,
E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
Synthesis of [11C]interleukin 8 using a cell-free translation system and L-[11C]methionine.
January 2012, Nuclear medicine and biology,
E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
Synthesis of functionally active human proinsulin in a cell-free translation system.
January 2005, Doklady. Biochemistry and biophysics,
E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
Continuous-flow cell-free translation, transcription-translation, and replication-translation systems.
January 1998, Methods in molecular biology (Clifton, N.J.),
E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
Cell-free translation of a biologically active, antigen-specific suppressor T cell factor.
June 1982, Proceedings of the National Academy of Sciences of the United States of America,
E V Volyanik, and A Dalley, and I A McKay, and I Leigh, and N S Williams, and S A Bustin
Production of dihydrofolate reductase by an improved continuous flow cell-free translation system using wheat germ extract.
June 1993, The Tokushima journal of experimental medicine,
Copied contents to your clipboard!