Use of flow field-flow fractionation for the rapid quantitation of ribosome and ribosomal subunits in Escherichia coli at different protein production conditions
✍ Scribed by Mikael Nilsson; Leif Bülow; Karl-Gustav Wahlund
- Publisher
- John Wiley and Sons
- Year
- 1997
- Tongue
- English
- Weight
- 200 KB
- Volume
- 54
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
Asymmetrical flow field-flow fractionation was and subunits due to the rather laborious experiments used for rapid (8-14 min) separation of ribosomes and and the experimental skill required (Birnbaum and their subunits. The amount of ribosomes and the mass Bailey, 1991). Moreover, artifacts occurring during ulfraction of ribosomes was determined in growing Eschetracentrifugation have been reported (Hauge, 1971; richia coli cells. These quantities changed significantly Spirin, 1971). These are connected to factors such as at different growth phases. Ribosomal composition was monitored after the insertion of a protein-encoding plas-the long process time and the high hydrostatic pressure.
mid and after the addition of an antibiotic agent. The
Improvements in the analytical methodology can open
results suggest that the method will be useful in studies up new possibilities to study ribosomal systems.
of, e.g., the relationships between the protein production
We have previously shown (Nilsson et al., 1996) that capacity of cells and the ribosomal composition. The analasymmetrical flow field-flow fractionation (asymmetriysis time is substantially shorter than ultracentrifugation run times.