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Two PZAase-negative mutants out of 4,000 colonies screened were identified.
AnpyrG insertion into the second intron of Aaku80 successfully disrupted Aaku80 function, which was restored by intramolecular HR between 20-bp DRs. Furthermore, the coku80 strain stably maintained the AnpyrG gene after 10 generations and yielded 17 xylan-utilization-deficient mutants out of 21 transformants (81%).
This initial screening step identified 6 nontoxic mutants out of the total of 28 derivatives.
Only three mutants, out of the eleven investigated, show a significant increase in the initial oligomerization rates compared to the WT protein.
At weaning, only eight Skm2 mutants out of 84 progeny were identified (9.5% ratio), suggesting embryonic or perinatal mortality.
Typically, 10 15 mutants out of ~20,000 were selected for further validation during each round of screening.
Similar(46)
Consistently, more mutants (3 out of 46 PCR clones) were detected by DNA sequencing.
We show that proper blood vessel pathfinding requires the endothelial receptor PlexinD1 and semaphorin signals, and we identify mutations in plexinD1 in the zebrafish vascular patterning mutant out of bounds.
The mutant simulation experiments show realistic results, and only one mutant (out of five) could not fully be reproduced.
One mutant out of the 3600 screened showed dark red colony color on low adenine medium plate and was identified as an ade7 - insertion mutant (Table 2 and mutant 13_C10 in Additional file 2: Table S1).
Expression of >400 genes was shown to be upregulated in the Salmonella hns mutant, out of which more than 90% were acquired by the horizontal gene transfer (Navarre et al., 2006).
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