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Gene information
General information
TC003474 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_007418.3: 5667930..5670294
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
    TC003474
    iBeetle screen iB_00573

    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)

    pupa eclosion not fulfilled (died as pupa) - 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


    Fertility
    mature egg deposition 4 number 2nd lateral oviduct: number 1st lateral oviduct: blocked - Number of animals showing the phenotype: 3
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 20
    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: 60.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection


    pupal molt not fulfilled (died as pupa) - Number of animals showing the phenotype: 5 (11 dpi)
    Days post injection


    pupal molt delayed (still larva) - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection


    larva lethality late (L7) - Number of animals showing the phenotype: 1 (16 dpi)
    Days post injection


    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 2 (16 dpi)
    Days post injection


    Pupal morphology
    wings blistered - Number of animals showing the phenotype: 6

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC003474 RA
      MDEEYDERN WDTVDRLDN DSDAGDETE NPEQVLKEC 
      AEKFSTSDY IMEPGIFSQ LKRYFQSGG NPLQVIEEL 
      SQNYTAVAQ MANLIAEWL ITGGVNVTS VQAMVENHL 
      KEMILKTFD PKKADTIFT EEGETPAWL TQLIEHPTW 
      RSLIYRLAE EYPDCLMLN FTIKLISDA GFQAEITSI 
      STAAQQLEV FSRVLKTSI ASFLTNPED WQNTSKECA 
      KMVCHGQHT YVYSQVIIH VLARESKGG SSMKRLSQE 
      ITKCAQKNG HDVTPITMA LNGASAYPL ACQALTSML 
      SKNTLNPAD ITVLYRNYN APDPPPIDL IRTPQFLEL 
      LIDALFKPG VKLNPDHKP KYIHLLAYA ASVYEMQAK 
      KGQKRVVNK DELQATIHS IETVHNICN TNKGSSELM 
      AELATLYQC LRYPVVSVG IIRWVECTV TEASYFKLS 
      TEHTPIHLA LLDEVVTCH PLLHNQVLD LLIRLFESK 
      QDELEILVQ LEMRKMVLD RMVNLLCAG CVVPVVKYI 
      KQCWQKGDT DISLIRYFV TEVLETIGP PYSSEFVHL 
      FMPMVENDE ITGTMRGDG ENDPVSEFI VYCKANYTA 
      VV
    CDS sequence
    • >TC003474 RA
      atggatgaa gaatacgac gagagaaat tgggatacc 
      gttgatcga ctcgataat gactcagat gcgggcgat 
      gaaacggaa aaccctgag caggtcctt aaggaatgt 
      gcggagaaa ttttcgacc tcagattac ataatggaa 
      ccgggaatt ttctcccag ctgaaacgg tatttccag 
      tctggagga aaccccttg caagtgatt gaagaatta 
      tcacagaac tacaccgcc gtagcccag atggctaac 
      ttgatcgca gaatggcta atcacaggt ggagtcaat 
      gtaacttcg gttcaagca atggtggag aaccactta 
      aaagagatg attttgaaa acatttgat cccaaaaaa 
      gctgacacg attttcaca gaagagggt gagacgcca 
      gcttggctg acacagcta attgaacat cccacgtgg 
      cggtcactg atttataga ttagctgag gagtatcca 
      gactgctta atgttaaat ttcacaatt aaacttatt 
      tccgacgcc ggtttccaa gctgaaatc acaagcatt 
      tcaacggca gctcaacag ttagaagtc ttctctaga 
      gtactcaaa acttcaatt gctagtttt ttgaccaat 
      ccagaagac tggcaaaat accagcaag gagtgtgca 
      aaaatggtg tgccacggc caacacacg tacgtttac 
      agtcaagtg attatccac gtgttagct cgagaatcc 
      aagggtggc agtagcatg aagcgactg tctcaagaa 
      atcacgaaa tgcgcccaa aaaaacggt catgacgta 
      actccaatc acaatggcg ttaaacggc gcctctgca 
      taccctctg gcctgccaa gccttaaca tcaatgctt 
      tcaaaaaat acacttaat ccagccgac attacagtt 
      ctgtacaga aattacaac gctccagac cctcctccc 
      attgattta atcagaact ccgcaattt ctggaatta 
      ttgattgat gccttgttt aaacctgga gtaaaactc 
      aaccccgac cacaagccc aaatatatc catttgtta 
      gcatatgca gcaagtgtc tatgaaatg caagccaaa 
      aaagggcaa aaaagagtg gtcaataaa gatgaactt 
      caagcgact attcattca attgaaacg gtccacaat 
      atctgtaat accaataaa gggagttca gagctaatg 
      gcagaactt gcaacgctg tatcagtgt ctcaggtat 
      ccagtcgtt tctgttgga attatccgt tgggttgag 
      tgcactgtt accgaagca agttacttt aaattgtca 
      acagaacac actccaatt catttggcg cttttggat 
      gaagtagtt acttgtcac cctctgctc cataatcaa 
      gtactcgac ttactaatc aggttgttt gaatccaag 
      caagacgaa ctggaaatt ttagtccaa cttgaaatg 
      cgtaaaatg gtccttgac agaatggta aatttgcta 
      tgtgctgga tgtgtggta cccgttgta aagtacata 
      aaacagtgt tggcaaaaa ggcgacacg gacatatcc 
      ttgattcgt tactttgta acagaagtt ctggagacc 
      atagggccc ccttatagc tccgaattc gtccactta 
      tttatgcca atggtcgaa aatgacgaa attacggga 
      acaatgaga ggagatggg gagaacgac cccgtgtca 
      gaatttatc gtctattgc aaagcaaat tacaccgca 
      gttgtataa
    mRNA sequence
    • >TC003474 RA
      cagtaacgt caaatttcg aaggttatg ttatggtta 
      aggacgcga ctcttttta ttgttttaa tttaactgc 
      cccaattat atgtaatta acttgcaat ggatgaaga 
      atacgacga gagaaattg ggataccgt tgatcgact 
      cgataatga ctcagatgc gggcgatga aacggaaaa 
      ccctgagca ggtccttaa ggaatgtgc ggagaaatt 
      ttcgacctc agattacat aatggaacc gggaatttt 
      ctcccagct gaaacggta tttccagtc tggaggaaa 
      ccccttgca agtgattga agaattatc acagaacta 
      caccgccgt agcccagat ggctaactt gatcgcaga 
      atggctaat cacaggtgg agtcaatgt aacttcggt 
      tcaagcaat ggtggagaa ccacttaaa agagatgat 
      tttgaaaac atttgatcc caaaaaagc tgacacgat 
      tttcacaga agagggtga gacgccagc ttggctgac 
      acagctaat tgaacatcc cacgtggcg gtcactgat 
      ttatagatt agctgagga gtatccaga ctgcttaat 
      gttaaattt cacaattaa acttatttc cgacgccgg 
      tttccaagc tgaaatcac aagcatttc aacggcagc 
      tcaacagtt agaagtctt ctctagagt actcaaaac 
      ttcaattgc tagtttttt gaccaatcc agaagactg 
      gcaaaatac cagcaagga gtgtgcaaa aatggtgtg 
      ccacggcca acacacgta cgtttacag tcaagtgat 
      tatccacgt gttagctcg agaatccaa gggtggcag 
      tagcatgaa gcgactgtc tcaagaaat cacgaaatg 
      cgcccaaaa aaacggtca tgacgtaac tccaatcac 
      aatggcgtt aaacggcgc ctctgcata ccctctggc 
      ctgccaagc cttaacatc aatgctttc aaaaaatac 
      acttaatcc agccgacat tacagttct gtacagaaa 
      ttacaacgc tccagaccc tcctcccat tgatttaat 
      cagaactcc gcaatttct ggaattatt gattgatgc 
      cttgtttaa acctggagt aaaactcaa ccccgacca 
      caagcccaa atatatcca tttgttagc atatgcagc 
      aagtgtcta tgaaatgca agccaaaaa agggcaaaa 
      aagagtggt caataaaga tgaacttca agcgactat 
      tcattcaat tgaaacggt ccacaatat ctgtaatac 
      caataaagg gagttcaga gctaatggc agaacttgc 
      aacgctgta tcagtgtct caggtatcc agtcgtttc 
      tgttggaat tatccgttg ggttgagtg cactgttac 
      cgaagcaag ttactttaa attgtcaac agaacacac 
      tccaattca tttggcgct tttggatga agtagttac 
      ttgtcaccc tctgctcca taatcaagt actcgactt 
      actaatcag gttgtttga atccaagca agacgaact 
      ggaaatttt agtccaact tgaaatgcg taaaatggt 
      ccttgacag aatggtaaa tttgctatg tgctggatg 
      tgtggtacc cgttgtaaa gtacataaa acagtgttg 
      gcaaaaagg cgacacgga catatcctt gattcgtta 
      ctttgtaac agaagttct ggagaccat agggccccc 
      ttatagctc cgaattcgt ccacttatt tatgccaat 
      ggtcgaaaa tgacgaaat tacgggaac aatgagagg 
      agatgggga gaacgaccc cgtgtcaga atttatcgt 
      ctattgcaa agcaaatta caccgcagt tgtataaca 
      gcaacttaa taataagaa tgtctatta
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY