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Gene information
General information
TC004466 Gene

The sequence from the iB fragment is matched against the genes from the official gene set. The matche(s) is/are visualizied in a genome browser.

NC_007417.3: 4625075..4626568
Sequence information
Transcripts / Proteins (1) Get mRNA sequence
Get CDS sequence
Get protein sequence
Ortholog information (from OrthoDB)

The information about the homolog genes in Drosophila is provided by OrthoDB http://www.orthodb.org/. The homologs are sorted according to their similarity score. For an orthology statement additional analysis are required.
The data basis for the orthologs is OrthoDB v9 containing OGS3 for Tribolium castaneum and 2015_04 (r6.07) for Drosophila melanogaster.

Closest fly homologs
    TC004466
    iBeetle screen iB_00703

    Attention! iBeetle is a first pass screen. Hence, the documented phenotypes need to be confirmed by independent experiments using non-overlapping dsRNA fragments! Phenotypes with a penetrance > 50% are frequently reproduced. See Schmitt-Engel et al. 2015 for details on reproducibility.

    iB sequence
    Phenotype after pupal injection
    Usually 10 injected animals

    Pupal injection procedure

    Day 0: 10 female pupae of the pBA19 strain (muscle enhancer trap line) were injected with dsRNA.

    3 days post injection (dpi): Hatch control: Pupal and adult lethality as well as metamorphosis defects (molting, eclosion) were documented. For mating, 4 males of the black strain were added.

    9 dpi: First egg-lay was collected and incubated for cuticle analysis. Adult lethality and egg production (reduced/ no egg-lay) was documented.

    11 dpi: Second egg-lay was collected and incubated for embryonic muscle analysis. Adult lethality and egg production (reduced/ no egg-lay) were documented.

    Note: The adult morphology was not analyzed systematically at 3/9/11 dpi. Only obvious phenotypes, visible without magnification, have been annotated.

    13 dpi: Egg productivity and Ovary analysis
    The percentage of hatched larvae was documented and not hatched larvae/ eggs were embedded for cuticle analysis (15 dpi). In case of a reduction of egg production, 4 injected females were dissected to analyze the gross morphology of the ovaries.

    14 dpi: Analysis of embryonic musculature and early embryonic development
    Offspring of the injected females (hatched and not hatched larvae/ eggs) were analysed for embryonic lethality and muscle defects.

    15 dpi: Analysis of larval instar 1 cuticle
    Offspring of injected females were analysed and cuticle phenotypes were annotated.

    22 dpi: Stink gland analysis
    Documentation of defects in abdominal and thoracic stink glands (colour, size, content) of the injected femals.

    Terms used in the pupal injection procedure

    larval pantagmatic defects
    At least two tagmata (head, thorax, abdomen or terminus) show similar/ comparable defects.
    empty eggs
    Just empty shells are visible, no cuticle has been developed.
    strong defects
    L1 cuticle larvae which show severe defects. These strong defects are subdivided into three categories
    strongly fragmented (cuticle crumbs)
    No segments or appendages are identifiable, only residues of cuticle and bristles can be identified.
    cuticle remnants (with unidentifiable segments)
    Segments or parts of appendages are recognizable, but identity is not clear.
    cuticle remnants (with some identifiable segments)
    At least one segment or appendage is clearly regonizable.
    number of eggs on slide
    affected embryos usually (not always) do not hatch. Only the not hatched were used for cuticle analysis.
    For the embryonic muscle analysis the hatched as well as the not hatched were analysed.
    total number of affected eggs/embryos/larvae
    Summary of all different phenotypes/ phenotypic series which were annotated.
    inside-out
    The L1 larva shows a partial or complete inversion., e.g. bristles, appendages or parts of the abdominal segments are inverted into the interior of the cuticle.
    starved
    Dissected females show a strong resorption of fatbody predominately in the abdomen and the oogenesis is blocked. These are considered as eventually lethal phenotypes and the blocked oogenesis probably a secondary defect due to starvation.
    eclosion not fulfilled
    The emergence of the adult from the pupa stage is interrupted. This phenotype shows pupal as well as adult features.
    Please see the help page for more information
    Metamorphosis and survival

    Lethality 11 days after pupal injection: 0.0% (includes death as pupa, adult)

    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 embryo/egg not developed/not fertilized - Penetrance (percentage of animals/eggs on slide): less than 30%

    larval musculature fluorescence decreased - Penetrance (percentage of animals/eggs on slide): less than 30%

    larval musculature random pattern irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    remarks: muscle pattern potentially obscured by segmentation defect,
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): more than 80%
    cuticle remnants with some identifiable segments - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    Phenotype after larval injection
    Usually 10 injected animals

    Larval injection procedure

    Day 0: 10 female L5-6 larvae of the D17Xhom strain (females express RFP in the eyes and in the CNS; pupae express GFP in the thorax) were injected with dsRNA.

    11 days post injection (dpi): Pupal morphology Documentation of larval/ pupal death and defects of metamorphosis (eclosion, delayed development). Screening for alterations of pupal head and appendages, pupal thorax, pupal thoracic musculature, pupal legs, pupal wings, pupal abdomen and pupal genital lobes.

    16 dpi: Adult morphology Documentation of larval/ pupal and adult death and defects of metamorphosis (eclosion, delayed development). Screening for alterations of adult head and appendages, adult thorax, adult elytra and hindwings, adult legs and cuticle properties. For mating 4 males of the black strain were added.

    19 dpi: Sieving and egg-lay Documentation of adult lethality and enduring metamorphosis defects.

    22 dpi: Ovary analysis (Fertility) In case of a reduction of egg production 4 females were dissected and the gross morphology of the ovaries were analysed.

    41 dpi: Fertility and stink gland analysis Documentation of defects in abdominal and thoracic stink glands development (colour, size, content) of the injected females. Check for fertility of injected larvae (no or decreased offspring).

    Terms used in the larval injection procedure

    pupal/ adult pantagmatic defects
    At least two tagmata (head, thorax, abdomen or terminus) show similar/ comparable defects.
    pupal molt not fulfilled
    The hatching from the larva to the pupa is interrupted. Either it dies as prepupa or the phenotype shows larval as well as pupal features.
    eclosion not fulfilled
    The emergence of the adult from the pupa stage is interrupted. This phenotype shows pupal as well as adult features.
    starved
    Dissected females show a strong resorption of fatbody predominately in the abdomen and the oogenesis is blocked. These are considered as eventually lethal phenotypes and the blocked oogenesis probably a secondary defect due to starvation.
    Metamorphosis and survival

    Lethalities 11 days after larval injection: % (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: % (includes death as larva, prepupa, pupa, adult)

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC004466 RA
      MECLSDTSD TSATSFERG RKPVDVKRP VLLVRYPKI 
      ELFPSRSDI ERLTAMGYG FTPEPAVFG LNWTIDQIN 
      EHIYSLYPD VPLQTIGFK MGKCDRTKK VFILKPVEN 
      TKELKEELG NSKLIILPN EELPERPIM HKKDEDSDE 
      DNTSLTRRR ERRTLNRYK LSEDDEDYE VIIENGQTP 
      KTKETNEEH EEYSVEKIL DKRTLNGKV EYLLKWKGY 
      SEQDNTWEP EENLDCPDL IAAYEAQFE VTPKADPEE 
      NEKDSKRKN ITEDNRARG FERGLEPER IVGATDASG 
      ELMFLMMWK NCKEADLVP ASQVSVRCP EVVINYYEN 
      KELWFCKQS VIEID
    CDS sequence
    • >TC004466 RA
      atggaatgc ttgtccgat acttccgac actagtgca 
      accagtttt gagcgaggc cggaagcca gtggatgta 
      aaaagacca gtcttgctt gtccgctac ccaaaaatc 
      gaattattt ccatctaga agtgatatt gagcgatta 
      actgctatg ggttatgga ttcactcca gaacctgca 
      gtgtttggc ctcaattgg acaattgat cagattaat 
      gaacacatc tattcgtta taccccgac gttcctttg 
      cagactatc ggtttcaaa atgggcaaa tgtgaccga 
      acaaaaaaa gtgttcatt ctaaaacct gttgaaaat 
      acaaaagaa ctaaaagag gagctgggt aatagtaaa 
      ctaatcatt ctaccaaac gaggaactt cctgagaga 
      ccaattatg cacaagaag gatgaagac agtgatgag 
      gataacaca tctcttaca agaagaagg gagaggaga 
      actttaaac aggtacaag ttgagtgaa gatgacgag 
      gattacgag gtcataatt gagaatgga caaacccct 
      aaaacaaag gaaaccaac gaagaacat gaggagtat 
      tccgttgag aaaattctc gataagcgg acgcttaat 
      ggaaaagtc gagtatctt ctaaaatgg aagggatat 
      tccgaacaa gacaatact tgggaacct gaggagaat 
      ttggactgt cctgactta atagccgcc tatgaggcg 
      cagtttgaa gtcacacct aaagcggac cctgaagag 
      aatgaaaaa gacagtaaa agaaaaaat attacggag 
      gataatcgg gcgcgcgga tttgagcgt ggcctcgag 
      cctgaaagg attgttggt gctactgat gccagtgga 
      gagcttatg tttctaatg atgtggaaa aattgcaaa 
      gaagcggat ttggtacca gcgagtcag gtcagcgtg 
      aggtgtccc gaggtcgtt attaattat tacgagaat 
      aaagaattg tggttttgt aaacagagc gtcatcgag 
      attgattaa
    mRNA sequence
    • >TC004466 RA
      gcgcaacaa ttataattg ttttgattt aagtttaat 
      cgtttaatt aagtttata aaagtcgta aattatcaa 
      aaacgttcc taagtgtaa taattatgg ttactttat 
      accacagaa aaggcccag aataacttc attatagat 
      ctgattaat tttttacag acgcaaagt ttacataac 
      ctcacaaaa atggaatgc ttgtccgat acttccgac 
      actagtgca accagtttt gagcgaggc cggaagcca 
      gtggatgta aaaagacca gtcttgctt gtccgctac 
      ccaaaaatc gaattattt ccatctaga agtgatatt 
      gagcgatta actgctatg ggttatgga ttcactcca 
      gaacctgca gtgtttggc ctcaattgg acaattgat 
      cagattaat gaacacatc tattcgtta taccccgac 
      gttcctttg cagactatc ggtttcaaa atgggcaaa 
      tgtgaccga acaaaaaaa gtgttcatt ctaaaacct 
      gttgaaaat acaaaagaa ctaaaagag gagctgggt 
      aatagtaaa ctaatcatt ctaccaaac gaggaactt 
      cctgagaga ccaattatg cacaagaag gatgaagac 
      agtgatgag gataacaca tctcttaca agaagaagg 
      gagaggaga actttaaac aggtacaag ttgagtgaa 
      gatgacgag gattacgag gtcataatt gagaatgga 
      caaacccct aaaacaaag gaaaccaac gaagaacat 
      gaggagtat tccgttgag aaaattctc gataagcgg 
      acgcttaat ggaaaagtc gagtatctt ctaaaatgg 
      aagggatat tccgaacaa gacaatact tgggaacct 
      gaggagaat ttggactgt cctgactta atagccgcc 
      tatgaggcg cagtttgaa gtcacacct aaagcggac 
      cctgaagag aatgaaaaa gacagtaaa agaaaaaat 
      attacggag gataatcgg gcgcgcgga tttgagcgt 
      ggcctcgag cctgaaagg attgttggt gctactgat 
      gccagtgga gagcttatg tttctaatg atgtggaaa 
      aattgcaaa gaagcggat ttggtacca gcgagtcag 
      gtcagcgtg aggtgtccc gaggtcgtt attaattat 
      tacgagaat aaagaattg tggttttgt aaacagagc 
      gtcatcgag attgattaa aacaaataa agtagtatt 
      tcgtaa
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY