If the transformant colonies on MacConkey plates were all pale-coloured, the ZFR was deemed to be a recombination-proficient mutant, so the expression plasmid was isolated and the ZFR reading frame was sequenced.
However, no difference in the internalisation rate of total cell-surface α6 integrin was observed in cells overexpressing the CD151 YXXΦ mutant, so the magnitude of its impact on integrin trafficking remains to be determined.
For H3K27me3, no significant difference was found at STM between the wild-type and the sdg8 mutant, so STM was used as the reference gene.
Given that there was no significant loss of a class of more severely affected animals, we suggest that the apparent difference reflects a delay in tissue replacement in the mutant, so that most animals end up with milder defects by the end of pupariation than were apparent during the time window for making the movies.
Given that there was no evidence for loss of a class of more severely affected animals, we suggest that the apparent difference reflects a delay in tissue replacement in the mutant, so that most animals end up with milder defects by the end of pupariation than were apparent during the early pupal time-window in the videos.
A Southern blot analysis revealed that the T-DNA was inserted into a single locus in the alb1 mutant, so we performed TAIL-PCR to identify the T-DNA flanking sequences.
A similar pattern of CO protein accumulation was observed in spa1 7 and cop1 4 mutants between 14 h after dawn and the following morning, but the rise in the abundance of the protein was delayed in the spa1 7 mutant, so that it could not be detected until 14 h after dawn.
This is consistent with the idea that the Aux/IAA-ARF network is differently configured but not globally down-regulated in the mutant, so that some phenotypes are suggestive of auxin resistance, but others are not.
As in all genetic screens, primary screens represent a rough first pass through a large number of mutants so that the bulk of the analysis (secondary screens) can be focused on the most promising candidates.
In addition, the proposed debugging technique can avoid the computation of unnecessary mutants so that the results show 2.9× average reduction of the number of mutants to be processed.
We were not, however, able to isolate the N-terminal deleted mutant, so are unable to comment on its ability to interact with XPO2.
