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Gene information
General information
TC003996 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: 28927334..28929485
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
    TC003996
    iBeetle screen iB_03666

    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)

    Fertility
    mature egg deposition >5 number 2nd lateral oviduct: number 1st lateral oviduct: blocked - Number of animals showing the phenotype: 4
    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: < 20 embryo/egg with embryonic tissue, no cuticle, but muscle fluorescence - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    embryo/egg with embryonic tissue potentially present, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): more than 80%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - 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: 20.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 90.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality - Number of animals showing the phenotype: 2 (11 dpi)
    Days post injection


    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 4 (16 dpi)
    Days post injection



    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 3 (16 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 1 (41 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC003996 RA
      MESNKDEAE KCIKLAETY IKERNREKA EKFLHKAER 
      LYPSQKAQD LLIQIKLMA ETKAETEQP RKRNVAPRT 
      EEVKKAPEY TQEQLEAVK KINKCKDFY EILGVSKDA 
      TDSEIKKAY KKLALQFHP DKNKCPGAA EAFKKIGNA 
      VAILTDVEK RKQYDQFGP EDERLSTRT HYSNTFREF 
      DADATDEIF NMFFGGGFN GANVYVRRG GRWQRQNAG 
      SQETHHNHH HREQNNYSA FVQLLPIIL AIVLSMASS 
      FFVSDPPYS LQAHAKYPI ARQTANLKI PYYVKENFA 
      TEFQGSVRR LEMSVEEEY VSNLRHACY REKNYRDSM 
      IWKARNFGD KELFQAAQN IKTPACEKL QTLR
    CDS sequence
    • >TC003996 RA
      atggaaagc aacaaagac gaagctgaa aagtgcatc 
      aaactagcc gaaacatac atcaaagaa cgtaaccgc 
      gaaaaagcc gagaaattt cttcacaaa gctgaacgt 
      ctctatcct tcacaaaaa gcccaagat ttactcatc 
      caaatcaaa ctcatggcc gaaaccaaa gctgagacc 
      gaacaaccg agaaagcgc aacgttgct cccagaaca 
      gaggaggtc aaaaaggcc cccgagtac acgcaagag 
      cagctcgaa gccgtcaag aaaatcaac aaatgcaag 
      gacttttac gagattttg ggggttagc aaagacgcg 
      accgattcg gaaatcaag aaggcgtac aaaaaacta 
      gccttgcag ttccatcct gacaagaac aagtgtcct 
      ggggccgca gaagccttc aaaaaaatt ggcaacgct 
      gtggcgatt ttaaccgat gttgaaaaa cgcaaacaa 
      tacgaccag tttggccct gaagatgaa cgtctttcg 
      acgagaact cactacagt aacacattt agggaattt 
      gatgccgac gcaactgat gaaattttc aatatgttc 
      tttggtggt ggttttaac ggggctaat gtctacgta 
      agaagaggg ggaaggtgg caaagacag aatgctgga 
      tctcaggaa acccatcac aatcatcat cacagagaa 
      caaaataac tactcggca tttgtgcag cttttgcca 
      atcattttg gcaattgta ttgtccatg gcgtctagt 
      ttcttcgtt tctgatccg ccatacagt ctacaagcg 
      catgcgaaa tacccaatc gcacgacaa acagcaaat 
      ttaaaaatc ccatactat gttaaagaa aatttcgca 
      acggagttt caagggtca gttcgaagg cttgaaatg 
      tcagtggag gaggagtat gtctcgaat ttaagacac 
      gcttgttac agagagaaa aattatcgt gattcgatg 
      atttggaag gctaggaat tttggagac aaggaactg 
      tttcaggcc gctcaaaat ataaaaacg ccagcgtgt 
      gaaaaactt caaacgttg cgttaa
    mRNA sequence
    • >TC003996 RA
      gtcaaaatc atctgtttt cccttcaat taaccatca 
      caaaaaaat caattaaat cgaaggttt ctcgtctcc 
      gacactaat atctcacga tggaaagca acaaagacg 
      aagctgaaa agtgcatca aactagccg aaacataca 
      tcaaagaac gtaaccgcg aaaaagccg agaaatttc 
      ttcacaaag ctgaacgtc tctatcctt cacaaaaag 
      cccaagatt tactcatcc aaatcaaac tcatggccg 
      aaaccaaag ctgagaccg aacaaccga gaaagcgca 
      acgttgctc ccagaacag aggaggtca aaaaggccc 
      ccgagtaca cgcaagagc agctcgaag ccgtcaaga 
      aaatcaaca aatgcaagg acttttacg agattttgg 
      gggttagca aagacgcga ccgattcgg aaatcaaga 
      aggcgtaca aaaaactag ccttgcagt tccatcctg 
      acaagaaca agtgtcctg gggccgcag aagccttca 
      aaaaaattg gcaacgctg tggcgattt taaccgatg 
      ttgaaaaac gcaaacaat acgaccagt ttggccctg 
      aagatgaac gtctttcga cgagaactc actacagta 
      acacattta gggaatttg atgccgacg caactgatg 
      aaattttca atatgttct ttggtggtg gttttaacg 
      gggctaatg tctacgtaa gaagagggg gaaggtggc 
      aaagacaga atgctggat ctcaggaaa cccatcaca 
      atcatcatc acagagaac aaaataact actcggcat 
      ttgtgcagc ttttgccaa tcattttgg caattgtat 
      tgtccatgg cgtctagtt tcttcgttt ctgatccgc 
      catacagtc tacaagcgc atgcgaaat acccaatcg 
      cacgacaaa cagcaaatt taaaaatcc catactatg 
      ttaaagaaa atttcgcaa cggagtttc aagggtcag 
      ttcgaaggc ttgaaatgt cagtggagg aggagtatg 
      tctcgaatt taagacacg cttgttaca gagagaaaa 
      attatcgtg attcgatga tttggaagg ctaggaatt 
      ttggagaca aggaactgt ttcaggccg ctcaaaata 
      taaaaacgc cagcgtgtg aaaaacttc aaacgttgc 
      gttaaaaaa atatttatt aatggaacg atcttgtta 
      tagttggag ttttgttgt tttattatc aacttgtat 
      tatttttgt tattatcta atatttatt gcaataagg 
      acctcacga tcattgtga tctattccc ggttagcaa 
      caaataaac acgatttta tgctacttg ccttattga 
      aatataaaa tttgtacac ttgtaccaa cccaaacgc 
      aaaatttgt accagaaat tttgcgtac gaagattac 
      caataaagt ggtttcttt aat
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY