The RMCE cassette of the Mi{MIC}brpMI02987 insertion has been replaced with a EGFP-FlAsH-StrepII-TEV-3xFlag multi-tag cassette. The insertion is in a coding intron of brp and is predicted to protein trap the gene.
brpMI02987-GFSTF.0 flies in which the level of brp protein has been reduced by expressing either slmbScer\UAS.T:Zzzz\vhhGFP4 (which targets GFP-tagged proteins for degradation) or an RNAi construct targeting GFP (which targets the GFP sequence-containing mRNA for degradation) under the control of Scer\GAL4ey.PU show electroretinogram (ERG) defects: there is a complete loss of both on- and off-transients, and ERG amplitude is reduced by 50% compared to controls.
brpMI02987-GFSTF.0 flies in which the level of brp protein has been reduced by expressing slmbScer\UAS.T:Zzzz\vhhGFP4 under the control of Scer\GAL4nSyb.PS and shifting to 28[o]C show a significant loss of excitatory junctional potential amplitude at the neuromuscular junction when the temperature shift occurs during the third larval instar. The severity of the defect becomes progressively more severe the earlier during the third larval instar that the shift is made.
The majority of brpMI02987-GFSTF.0 animals in which the level of brp protein has been reduced by expressing either slmbScer\UAS.T:Zzzz\vhhGFP4 or an RNAi construct which targets GFP sequence under the control of either Scer\GAL4nSyb.PS or Scer\GAL4da.PU continuously at 28[o]C show embryonic lethality.
brpMI02987-GFSTF.0 animals expressing either slmbScer\UAS.T:Zzzz\vhhGFP4 or an RNAi construct which targets GFP sequence under the control of Scer\GAL4nSyb.PS and shifted to 28[o]C from 18[o]C at 48 or 24 hours of development respectively show locomotor defects at the second larval instar stage and show late second instar lethality.
brpMI02987-GFSTF.0 animals expressing an RNAi construct which targets GFP sequence under the control of Scer\GAL4nSyb.PS and shifted to 28[o]C either at the second larval instar or adult stage show no obvious mutant phenotypes.
brpMI02987-GFSTF.0 animals expressing slmbScer\UAS.T:Zzzz\vhhGFP4 under the control of Scer\GAL4nSyb.PS and shifted to 28[o]C at the second instar stage show locomotor defects at the third larval instar stage and lethality at the pupal and adult stages.
brpMI02987-GFSTF.0 flies expressing slmbScer\UAS.T:Zzzz\vhhGFP4 under the control of Scer\GAL4nSyb.PS and shifted to 28[o]C after eclosion show locomotor defects and adult lethality.
