Biomedical Engineering Reference
In-Depth Information
R
ESULTS
The process of protein biosynthesis in procaryotic cells and the amount of the target
product are largely determined by cultivation conditions which should provide effective
expression of the cloned gene: realization of genetic and phenotypic properties of the
population of recombinant cells (identity and stability of plasmid, bacterial cell phenotype).
When cultivating
E.coli
JM 103/pTBI strain in YT medium, the effectiveness of expression
was low and the content of TBI protein in biomass was 5-8 % of the total protein. In order to
increase the yield of recombinant protein we optimized cultivation conditions of the
producing strain by varying the composition of the nutrient medium, the time of addition of
nalidixic acid and the aeration rate. At the first stage the process of microbiological synthesis
was carried out in flasks. TB medium with or without trace elements was chosen as an
experimental medium for cultivation of recombinant
E.coli
JM 103/p TBI strain. YT medium
served as a control. TB and TB plus media differ from YT medium by the multicomponent
composition and the presence of the additional source of carbon - glycerol [7-9].
Figure 1. The growth curves of
E.coli
JM 103/pTBI in optimized media.
Figure 2. The dynamics of accumulation of TBI protein in the process of cultivation of
E.coli
JM
103/pTBI in optimized media.
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