Inclusion body formation

Ralstonia eutropha, a Gram-negative bacterium that has the special feature of high-level expression without inclusion body formation, can also be regarded as a competitor to E. coli [59]. 

Two features of recombinant production in particular can impact very significantly upon the approach subsequently taken to purify the recombinant product inclusion body formation and the incorporation of purification tags. The processes of inclusion body formation, recovery and recombinant protein renaturation have been considered in Chapter 5. Once the recombinant protein has been refolded, additional purification (if required) follows traditional lines. 

Vesicular trafficking and inclusion body formation are both dependent on the integrity of microtubules and other cytoskeletal components. Parkin has been shown to target misfolded tubulin for degradation [251] (figure 4.6B) and to interact with centrosomes upon proteasomal inhibition [252]. Whether this reflects association with specific substrates or co-localization with proteasomes in centrosomes re-... 

La Vallie, E. et al. (1993). A thioredoxin gene fusion expression system that circumvents inclusion body formation in the E. coli cytoplasm. Bio j Technology 11, 187-193. 

In the publication by Nielsen et al. (2004) images of the pigmented inclusions that contain the phytoalexins were prepared by confocal microscopy. This provided a three-dimensional perspective of inclusion body formation and visualization of the phytoalexins. A representation of deoxyanthocyanidin accumulation is shown in Figure 6-3 where inclusions begin to form by 5 to 8 hours after a fungal appressorium was formed by a hypha. 

Model to Account for Competition between Folding and Inclusion Body Formation... 

Chrunyk, B. A., Evans, J., Lillquist, J., Young, P., and Wetzel, R. (1993) Inclusion body formation and protein stability in sequence variants of interleukin-1 beta. J. 

It is tempting to speculate if metabolic shifts and shifts in the physiological state of the culture can be more precisely monitored with the electronic nose. Physiologically important odorant metabolites in antibiotic strains such as geosmin and oxolone have been analyzed with conventional methods in the fermentation off-gas [38]. Recently, responses from electronic noses have been possible to relate to plasmid copy number [39]. Other types of intracellular shifts, such as viral infection, inclusion body formation or increase in mRNA level, could challenge the capacity of electronic noses. 

Another technique that Ernst et al.5 utilized was to reduce the temperature of the fermentor to 15°C during the induction phase. This reduced the inclusion body formation, and achieved soluble heparinase I at a level of 4.1 mg/g dry cell weight. Their calculations indicated that the manufacturing cost in this case would be 653,000 per kilogram. This is only 16% higher than the cost obtained for the insoluble heparinase I expression method. 

Masliah E, Rockenstein EM, Veinbergs I, Mallory M, Hashimoto M, Takeda A, Sagara Y, Sisk A, Mucke L (2000) Dopaminergic loss and inclusion body formation in a-synuclein mice implications for neurodegen-erative disorders. Science 287 1265-1269. 

Recently the rat 24-OHase cDNA was recombinantly overexpressed in a bacterial cell line and reconstituted with its required cofactors [225]. Cell fractionation revealed the 24-OHase to be primarily localized in membranes, with minimum inclusion body formation, and that upon reconstitution with adrenodoxin, adrenodoxin reductase, and NADPH, 24-OHase activity was apparent. Interestingly, this preparation produced three... 

Mitra S, Tsvetkov AS, Kinkbeiner S (2009) Protein turnover and inclusion body formation. Autophagy 5 1037-1038... 

Georgiou, G. and Bowden, G.A. (1991). Inclusion body formation and the recovery of aggregated recombinant proteins. In Recombinant DNA Technology and Applications. A.Prokop, R.K.Bajpai, and C.Ho, eds. (New York McGraw-Hill), pp. 333 356. 

Rinas, U., Tsai, L.B., Lyons, D., Fox, G.M., Stearns, G., Fieschko, J., Fenton, D., and Bailey, J.E. (1992). Cysteine to serine substitutions in basic fibroblast growth factor effect on inclusion body formation and proteolytic susceptibility during in vitro refolding. Bio/Technology 10,435 440. 

Chen Y, Song J, Sui SF, Wang DN. DnaK and Dual facilitated the folding process and reduced inclusion body formation of magnesium transporter CorA overexpressed in Escherichia coli. Protein Expr. Purif. 2003 32 221-231. 

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