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Gene information
General information
TC032269
Former TC numbers

If one gene is splitted or genes are merged, the original TC numbers are listed. One listed former TC number indicates the previous gene has be splitted, serveral former TC number represents a merged gene.

TC001364,TC001363
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: 15054901..15069523
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
    TC032269
    iBeetle screen iB_06859

    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: 50.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 2 (3 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 3 (9 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): more than 80%

    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
    cuticle remnants with unidentifyable segments - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    cuticle remnants with some identifiable segments - Penetrance (percentage of animals/eggs on slide): 50% - 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
    • >TC032269 RA
      MRSASAAAL LLAALLVVQ AVRACGPGR GVGRRRGPR 
      KLTPLVFKQ HVPNVPENT LTASGLTEG RIGRNDSRF 
      KDLVPNYNQ DIVFKDEEG TGADRLMTQ RCKEKLNTL 
      AISVMNQWP GVRLLVTEG WDEEGYHTP ESLHYEGRA 
      VDITTSDRD RSKYGMLAR LAVEAGFDW VYYESRAHI 
      HCSVKSESS QAAKYGGCF SGESTVLTS TGLRRNLSS 
      LQIGEKIQA LDPSTNELV FSEVLLFLD YNPSQRRQF 
      LHITLASGR TLTVTPSHL LVLDDRTMK YAQKLQPGD 
      FLLVSDNAK NALISEKIV RLEAVWRSG VFAPLTGVG 
      TLVVNDVVA SCYATIDSQ WLAHWAFAP IRWVAKLWD 
      SGLRKPGVG VFWYARLLY ATADFVLPS HLLHE
    CDS sequence
    • >TC032269 RA
      atgaggtct gcttcggca gcggcgctc ctgctagcg 
      gccctcctg gtggtgcag gcggtgcgg gcttgcggc 
      ccggggcgc ggagtcggc cggagacga gggccaagg 
      aagctcaca ccgctcgtc ttcaaacag catgtgccc 
      aatgtgccg gagaatacg ctcacggcg agcggactc 
      actgaagga cgcatcgga aggaatgac tccaggttc 
      aaggatctg gtgcccaat tataaccag gacatcgtc 
      ttcaaggat gaggagggc acgggggcg gacaggctc 
      atgacacag agatgtaag gagaagttg aacactttg 
      gcgatttcg gtgatgaac caatggccc ggtgttcgc 
      cttctggtg actgaaggc tgggatgaa gaaggctat 
      catacacca gagtccctt cactacgaa ggacgtgct 
      gttgatatc accaccagc gatcgagat cgctcgaaa 
      tacggaatg ttggccaga ctggctgtg gaagcgggc 
      tttgattgg gtctactat gaaagtagg gcccacata 
      cactgctcg gtcaaatca gaatcctcc caagccgca 
      aaatacggc ggttgtttc tcgggcgaa agcacagtc 
      ctgacctcc accggcctc cgtcgcaac ctctcctcg 
      ctccaaatc ggtgagaaa atccaagca ctggacccc 
      tccaccaac gagctcgtc ttcagtgag gtcctgctt 
      ttcctcgac tacaacccc tctcaaagg cgccaattt 
      ctccacata accctggct tcaggtcgc actttgaca 
      gtgacccca tctcatttg ctcgtcctc gacgatcgg 
      acaatgaaa tacgcgcaa aagctccaa ccgggcgac 
      tttttactc gtgagtgac aacgcgaaa aacgctcta 
      attagcgaa aaaatcgtg cgtttggag gccgtgtgg 
      cggagtggc gttttcgcg ccgttgacg ggcgtcggg 
      acgctggtg gtgaatgat gtggtggcg tcgtgttac 
      gcgaccatt gacagtcaa tggttggcc cattgggcg 
      tttgcgccc attagatgg gtggcaaag ttgtgggat 
      tccgggtta aggaagccg ggagttggg gttttttgg 
      tacgcgagg ttgctttat gctacggcg gactttgtt 
      ctaccttcg catttactg cacgagtga
    mRNA sequence
    • >TC032269 RA
      gcgtgcacc agtcattga aatctccag ctggtgatg 
      ttagttagt tagcagttg agttggtgg caggccaga 
      aatatagta tgatttgcg aggtgcagg ttcatctgc 
      ctcggagtt gggctccgc ccgctggcc aatagccgc 
      tgaggccgt ccgaagatg gccagggcc ttcaagtga 
      gtggggcca gtcagtcgg tgtttcacc tatgacaag 
      tgcaagatg aggagtgct gatggtgcg tgcgttcgg 
      agatgaggt ctgcttcgg cagcggcgc tcctgctag 
      cggccctcc tggtggtgc aggcggtgc gggcttgcg 
      gcccggggc gcggagtcg gccggagac gagggccaa 
      ggaagctca caccgctcg tcttcaaac agcatgtgc 
      ccaatgtgc cggagaata cgctcacgg cgagcggac 
      tcactgaag gacgcatcg gaaggaatg actccaggt 
      tcaaggatc tggtgccca attataacc aggacatcg 
      tcttcaagg atgaggagg gcacggggg cggacaggc 
      tcatgacac agagatgta aggagaagt tgaacactt 
      tggcgattt cggtgatga accaatggc ccggtgttc 
      gccttctgg tgactgaag gctgggatg aagaaggct 
      atcatacac cagagtccc ttcactacg aaggacgtg 
      ctgttgata tcaccacca gcgatcgag atcgctcga 
      aatacggaa tgttggcca gactggctg tggaagcgg 
      gctttgatt gggtctact atgaaagta gggcccaca 
      tacactgct cggtcaaat cagaatcct cccaagccg 
      caaaatacg gcggttgtt tctcgggcg aaagcacag 
      tcctgacct ccaccggcc tccgtcgca acctctcct 
      cgctccaaa tcggtgaga aaatccaag cactggacc 
      cctccacca acgagctcg tcttcagtg aggtcctgc 
      ttttcctcg actacaacc cctctcaaa ggcgccaat 
      ttctccaca taaccctgg cttcaggtc gcactttga 
      cagtgaccc catctcatt tgctcgtcc tcgacgatc 
      ggacaatga aatacgcgc aaaagctcc aaccgggcg 
      actttttac tcgtgagtg acaacgcga aaaacgctc 
      taattagcg aaaaaatcg tgcgtttgg aggccgtgt 
      ggcggagtg gcgttttcg cgccgttga cgggcgtcg 
      ggacgctgg tggtgaatg atgtggtgg cgtcgtgtt 
      acgcgacca ttgacagtc aatggttgg cccattggg 
      cgtttgcgc ccattagat gggtggcaa agttgtggg 
      attccgggt taaggaagc cgggagttg gggtttttt 
      ggtacgcga ggttgcttt atgctacgg cggactttg 
      ttctacctt cgcatttac tgcacgagt gatatgtat 
      aaaaaatta ggtttaggt tttttggcc agtgactta 
      ggtagtgcg gatagacct ttgagtgtt ccttaatta 
      attaattaa ttaatttat gacttgact tttttgtga 
      caaaaaact gtatgtagt gtatttttc atgtatata 
      caaaactac ttttacata ttgtatagt ttcttattg 
      tttgaaagt tgtacatag ttatttatg taatatata 
      gtccacttt tatttatgg tgagaaatt ttatatcgg 
      agacaaaat aaacgttaa tgtgatgc
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY