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The generated mutants were checked by sequencing and also by blotting against anti-GST antibodies (GE Healthcare Life Sciences).
Five replicates of TAF23 mutants were checked for the stability of G418 resistance gene, however they were not able to resist and to grow at the concentration 250 µg/mL of G418.
All mutants were checked using PCR amplification and sequencing.
At each step, mutants were checked with NuSMV, using the CTL properties above, but with a synchronous update rule.
Both the wild type dephosphocoenzyme A kinase and all the mutants were checked for their susceptibility to tryptic digestion by checking their fragmentation patterns upon exposure to trypsin under limiting conditions at a protease to protein ratio of 1∶50 (w/w).
All mutants were checked by DNA sequencing.
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For the determination of protein expression of ply, the expression of WT PLY from IDM- ply mutants was checked by blotting of rabbit polyclonal pneumolysin antibody (abcam, ab71811, Cambridge, UK).
Growth profiles of wild-type (WT) and the recC, recB, recD and recCBD null-mutants were checked by growing the strains in liquid broth at 22° and 4°C.
To check if this is metabolic or signaling effect of glucose, the effect of increasing concentration of glucose on gin2 (glucose receptor mutant) [6] mutant was checked.
The integrity of the folded state of each mutant was checked carefully, using CD and in some cases NMR spectroscopy.
The nucleotide sequences of the mutant cDNA were checked by DNA sequencing (Applied Biosystems).
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