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Gene information
General information
TC012574 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_007424.3: 11632168..11633272
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
    TC012574
    iBeetle screen iB_05273

    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): more than 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    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%
    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
    • >TC012574 RA
      MNFYVFVLC TLFASCYSK PNNVGIIGG KVARVGEFP 
      WQAAVYVDT ADGKFFCGG SLINNQWIL TAGHCVYNA 
      RLFTIHLGS ITLQAGDPN RVVLSTSTS VAFPNFDPL 
      TLEHDIGLI KLHMKVELS DYIQPIELA DIGAISEGS 
      AAIISGWGQ TADATAGLS NNLQYTNVV VISNNECKQ 
      TYGDQIKNT MICTAGNFN EGICTGDTG GPLIRAKGA 
      DNYVQIAVA GFFSQSGCE QLHPSGYIR TDLYKAWIQ 
      NVTETM
    CDS sequence
    • >TC012574 RA
      atgaatttc tacgttttt gttttgtgc acgcttttc 
      gcctcgtgt tattccaaa cccaacaat gttgggata 
      atcgggggg aaagtggcc cgcgttggg gaattcccc 
      tggcaagcg gccgtttac gtggacacc gctgacggc 
      aaattcttt tgtgggggg tctctgatc aacaatcag 
      tggattctg actgctggt cattgcgtt tacaatgcg 
      agacttttt acaattcat ttgggttca atcaccctg 
      caagctggg gaccccaac cgagtggtt ctctcaacc 
      tccacgtca gtcgctttc cccaatttc gaccccctg 
      actttggag cacgacatc gggcttatt aaactgcac 
      atgaaagtg gaactttct gattacata caaccaatc 
      gaactggcc gacattggg gccatttcc gagggcagt 
      gccgccata atctcagga tggggccaa accgctgac 
      gcaactgcc ggtttgtct aacaatctc caatacaca 
      aacgtggta gttatatca aacaatgag tgtaaacag 
      acctacggt gaccaaatc aaaaacacg atgatttgt 
      actgctggt aacttcaat gagggcatt tgcactgga 
      gacactggt ggtccgctg atccgagcc aaaggggct 
      gacaattat gtacaaatt gccgttgct ggatttttc 
      agtcagagt ggctgcgaa cagttgcac ccttcggga 
      tatatcagg accgatctg tacaaagcc tggattcag 
      aatgtaacc gaaacgatg tag
    mRNA sequence
    • >TC012574 RA
      gtccgctga taattcgga ttgttgaat accagtcat 
      gaatttcta cgtttttgt tttgtgcac gcttttcgc 
      ctcgtgtta ttccaaacc caacaatgt tgggataat 
      cggggggaa agtggcccg cgttgggga attcccctg 
      gcaagcggc cgtttacgt ggacaccgc tgacggcaa 
      attcttttg tggggggtc tctgatcaa caatcagtg 
      gattctgac tgctggtca ttgcgttta caatgcgag 
      actttttac aattcattt gggttcaat caccctgca 
      agctgggga ccccaaccg agtggttct ctcaacctc 
      cacgtcagt cgctttccc caatttcga ccccctgac 
      tttggagca cgacatcgg gcttattaa actgcacat 
      gaaagtgga actttctga ttacataca accaatcga 
      actggccga cattggggc catttccga gggcagtgc 
      cgccataat ctcaggatg gggccaaac cgctgacgc 
      aactgccgg tttgtctaa caatctcca atacacaaa 
      cgtggtagt tatatcaaa caatgagtg taaacagac 
      ctacggtga ccaaatcaa aaacacgat gatttgtac 
      tgctggtaa cttcaatga gggcatttg cactggaga 
      cactggtgg tccgctgat ccgagccaa aggggctga 
      caattatgt acaaattgc cgttgctgg atttttcag 
      tcagagtgg ctgcgaaca gttgcaccc ttcgggata 
      tatcaggac cgatctgta caaagcctg gattcagaa 
      tgtaaccga aacgatgta gaaattgtg attaataaa 
      ttgtttaaa acaaa
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY