General information
Escherichia coli (E. coli) is one of the most widely used hosts for the production of heterologous proteins and genetics are far better characterized than those of any other microorganism. A couple of strains with different genotypes are available for different, specific purposes, one of the work horses for recombinant protein expression being E. coli BL21.
Recent progress in the fundamental understanding of transcription, translation, and protein folding in E. coli, together with the availability of improved genetic tools like the IBA StarGate® technology are making this bacterium more valuable than ever for the expression of complex eukaryotic proteins when protein modifications like glycosylation are not a task.
Test Phase (please refer to Table 1)
- E. coli test culture (200 ml) and induction of protein expression
- Strep-Tactin® and/or Ni-NTA affinity purification, SDS-PAGE, Western blot
- Estimation of the culture volume required to produce the requested amount
of protein
- Time required: 1-2 weeks
Production Phase (please refer to Table 1)
- E. coli culture in the appropriate volume, preparation of protein extract and
Strep-Tactin® and/or Ni-NTA affinity purification
- Optimization
A set of different E. coli test cultures (200 ml) under various expression conditions and/or different affinity purification attempts can be performed. This set of experiments will be planned in close consultation with the client and depends on the initial results of the E. coli test scale expression.
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