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Practise questions minor 'Bioengineering and -omics'
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When should you use eukaryotic cells for gene expression?
When should you use eukaryotic cells for gene expression?
If you need post translational modification (yeast - correct protein folding), if you need glycosylation (animal cells), if you want to produce big proteins (>100 kD).
How can you modify 5' overhanging ends?
How can you modify 5' overhanging ends?
Use the Klenow Fragment of DNA polymerase I to fill in the overhang (5' to 3' DNA polymerase activity).
Remove overhangs by mung bean nuclease (cleaves ssDNA only).
How can you assay the activity of a protein in a cell?
How can you assay the activity of a protein in a cell?
Use a reporter gene (e.g., GFP, CAT, luciferase) to detect the amount of protein expressed by the promoter of the gene of interest. The reporter gene can be cloned after the to be studied promoter, after which the amount of detection marker can be easily assayed (visually or chemically).
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