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We mobilized some of the "revertant" leader sequences into our gene expression constructs and tested them for effects on ORF VII translation and found no or only marginal improvements of expression (data not shown).
More recently, the coexpression of protein disulfide isomerase, which probably reduces the retention of disulfide-bonded proteins in the endoplasmic reticulum, has led to enormous improvements of expression yields also in less dense cultures (Vad et al. 2005; Tsai et al. 2006; Huo et al. 2007).
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It is important to emphasize that synthetic gene engineering using Gene Composer does not always result in improved protein expression, and there was no clear improvement of expression of NS5B compared to native NS5B.
As evident from Figure 1, a minor improvement of expression yield could be detected.
Depending on the purpose of the application, there is still ample room for improvement of expression strategies, such as careful host (range) selection, co-expression of chaperones, or codon optimization.
However, activity recovery from inclusion bodies makes the whole process longer and more expensive, indicating that further improvements of the expression system and enzyme preparation protocol are required to make LipI.3 a faster and more efficient enzyme system.
Consequently, although the specific activity of both enzymes was improved successfully by indirect induction of chaperones or decelerated proliferation and expression rates, a further improvement of soluble expression was tested by coexpression of five different chaperone sets (see Additional file 1).
The entire situation is a litmus test for reform in our country, including for improvements in freedom of expression.
The 1.5 kb RNP CpG island fragment conferred improvements in level of expression from the hCMV promoter and percentage of cells showing detectable EGFP expression that were very similar to those observed for the larger fragments.
After improvement of the expression conditions, a hydantoinase and carbamoylase was successfully purified with two different methods.
The screening of experimental conditions for the improvement of enzyme expression coupled with the RSM experimental design serve as vital tool for analyzing the influence of cultivation conditions on the expression of recombinant bromelain.
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