Insect Mol Biol 9 : — The primary caveat when using these assays is their sensitivity to SNPs, which necessitates the use of uninjected sibling animals as negative controls. The output of this analysis provided the number of unique ZFN sites within each gene or the absence thereof for the 16, annotated unique genes within the genome.
Table 1. Butterfly genome reveals promiscuous exchange of mimicry adaptations among species. We verified that both mutations lead to the production of truncated proteins by Western blot analysis of brain tissue of wild-type, heterozygous, and homozygous mutants, using a monarch-specific anti-CRY2 antibody directed against the C terminus of the protein Fig.
Nat Biotechnol 25 : — CRY2 deficiency disrupts circadian behavior. Brain dissections were performed in 0. To assess the effect of monarch CRY2 knockouts on circadian behavior, we examined the time of day of adult eclosion the emergence of the adult butterfly from its chrysalis Fig. We recovered a single mutation at the targeted sequence transmitted by each founder, with one having a 4-bp deletion and the other a 2-bp insertion.
This analysis identified a high-quality ZFN site in exon 2 comprising two bp zinc-finger recognition elements flanking a 5-bp spacer containing an endogenous restriction site Fig. We found that the circadian timing of adult eclosion in DD was disrupted in both knockout lines, while sibling-matched wild-type and heterozygote animals exhibited normal eclosion timing Fig.
We used the EagI restriction site present in the spacer between the ZFN binding sites to detect the presence of lesions via the loss of sensitivity of polymerase chain reaction PCR —amplified fragments spanning this region to EagI digestion Fig. Science : J Insect Physiol 57 : — Preprint Server.
The molecular mechanism is unknown in both cases. One to two days prior to adult eclosion, the pupae were transferred to constant darkness DD.
Causal involvement of mammalian-type cryptochrome in the circadian cuticle deposition rhythm in the bean bug Riptortus pedestris. Proc Biol Sci : S — S Eclosion data were analyzed and plotted as 1-h bins. Predictably, we did not detect any signal at the expected size of full-length CRY2 in the brains of either knockout butterfly line Fig.