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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).
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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.
The mutants were checked by PCR and DNA sequencing.
Colonies were screened for replacement of the marker and point mutants were checked by sequencing.
The mutants were checked by sequencing using an ABI Prism 310 Genetic Analyzer (Applied Biosystems).
Mutants were checked for known growth defects http://www.broadinstitute.org/annotation/genome/neurospora/MultiHome.html.html
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