Status
Please wait ...
Gene information
General information
TC000771 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: 5702139..5703966
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
    TC000771
    iBeetle screen iB_00170

    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: 10.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 1 (3 dpi)
    Days post injection


    Fertility
    mature egg not present - Number of animals showing the phenotype: 3

    vitellogenic egg chamber potentially differentiation aborted - Number of animals showing the phenotype: 3

    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: < 50 embryo/egg with embryonic tissue, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    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: 37.5% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)

    pupal molt not fulfilled (died as L7 or prepupa) - 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: 2 (16 dpi)
    Days post injection



    adult lethality immediately after eclosion - Number of animals showing the phenotype: 1 (19 dpi)
    Days post injection


    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (19 dpi)
    Days post injection


    Pupal morphology
    wings shortened - Number of animals showing the phenotype: 2
    wings not closed - Number of animals showing the phenotype: 2

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC000771 RA
      MSLVNIVFR SRKSSLTKL RILKNLKKA RHFSVGALI 
      HQKNDEHIM HVFDRNTKS LQRERAASA QDANLYDYL 
      KDEIGYRLA DRVFDIKRK FKLAADIGC SRGYVSKHI 
      SPKCIEELI LCDMNRFNL DSVQVQEGI KVRKQVLDE 
      EHIEFEPNS LDLVISSLS LHWVNDLPN AFKQILKSL 
      KEDGVLLAA VFGGDTLYE LRSSLQLAE LERRGGISP 
      HISPFTEVR DIGNLLTRA GFTMLTIDT DEIVVNYPT 
      LFELMWDLK GMAESNAAI NRSLHLHRE TQFAAAAIY 
      QQLYGKTDP ETGKTTIPA TFQIINMLG WKPHPKQPK 
      PLERGSGEV SLKDLYKLD EIIKEVKKI KNDDDQKN
    CDS sequence
    • >TC000771 RA
      atgtctttg gttaatata gttttccgt tcacgtaaa 
      tcctctttg acaaaactc cgcattttg aagaatctt 
      aaaaaagcg agacacttt tcggtcggt gctttaatt 
      caccagaaa aatgacgaa cacattatg catgtgttt 
      gataggaac accaagagt ttgcaaagg gaacgggcg 
      gcttccgca caagacgcc aatttgtat gattatttg 
      aaagatgaa attggctat cgactagca gacagggtg 
      tttgatatc aaacgcaag tttaaactg gccgcagac 
      attggttgc agcaggggc tacgtgtcc aaacatatt 
      tcaccaaaa tgtattgaa gaactaatc ttatgtgat 
      atgaaccgc tttaattta gacagtgtt caagtacaa 
      gagggtatt aaagttagg aaacaagta ctagatgag 
      gaacacatt gagtttgag ccaaactcc cttgatctg 
      gttatatca agtttgagt ttgcattgg gtgaacgat 
      ttgccgaac gcttttaag caaatttta aaatcgctt 
      aaggaggat ggtgttctt cttgcggcg gtttttgga 
      ggagatact ttatatgag ttgagatct tccctccag 
      ttggctgaa ttagagcgt cgtggaggc atttctcct 
      cacatatct ccatttacc gaagttaga gatattggc 
      aatttgttg acaagggct ggcttcaca atgctcact 
      attgacact gatgaaatt gttgtgaat tatcctact 
      ttgtttgaa ttaatgtgg gaccttaaa ggaatggca 
      gagagtaat gctgctatt aataggtcg cttcatttg 
      caccgagag actcagttt gctgcagca gcaatttac 
      caacaactt tacggtaaa actgatcct gagacgggg 
      aaaaccaca atacctgcc acttttcaa atcattaat 
      atgttaggg tggaagccc catcccaaa caacctaaa 
      ccactagag aggggaagt ggggaagta tcattaaag 
      gatttgtac aaattggat gaaatcatt aaagaagta 
      aagaagatc aaaaacgac gatgatcag aagaattag
    mRNA sequence
    • >TC000771 RA
      catttgaca tttggttaa cctcaaaat cctaaactg 
      tcattgtga caattatga tttgttgtg ctgaaatga 
      gttgttttt aattcattt aaacagtgt catgtcttt 
      ggttaatat agttttccg ttcacgtaa atcctcttt 
      gacaaaact ccgcatttt gaagaatct taaaaaagc 
      gagacactt ttcggtcgg tgctttaat tcaccagaa 
      aaatgacga acacattat gcatgtgtt tgataggaa 
      caccaagag tttgcaaag ggaacgggc ggcttccgc 
      acaagacgc caatttgta tgattattt gaaagatga 
      aattggcta tcgactagc agacagggt gtttgatat 
      caaacgcaa gtttaaact ggccgcaga cattggttg 
      cagcagggg ctacgtgtc caaacatat ttcaccaaa 
      atgtattga agaactaat cttatgtga tatgaaccg 
      ctttaattt agacagtgt tcaagtaca agagggtat 
      taaagttag gaaacaagt actagatga ggaacacat 
      tgagtttga gccaaactc ccttgatct ggttatatc 
      aagtttgag tttgcattg ggtgaacga tttgccgaa 
      cgcttttaa gcaaatttt aaaatcgct taaggagga 
      tggtgttct tcttgcggc ggtttttgg aggagatac 
      tttatatga gttgagatc ttccctcca gttggctga 
      attagagcg tcgtggagg catttctcc tcacatatc 
      tccatttac cgaagttag agatattgg caatttgtt 
      gacaagggc tggcttcac aatgctcac tattgacac 
      tgatgaaat tgttgtgaa ttatcctac tttgtttga 
      attaatgtg ggaccttaa aggaatggc agagagtaa 
      tgctgctat taataggtc gcttcattt gcaccgaga 
      gactcagtt tgctgcagc agcaattta ccaacaact 
      ttacggtaa aactgatcc tgagacggg gaaaaccac 
      aatacctgc cacttttca aatcattaa tatgttagg 
      gtggaagcc ccatcccaa acaacctaa accactaga 
      gaggggaag tggggaagt atcattaaa ggatttgta 
      caaattgga tgaaatcat taaagaagt aaagaagat 
      caaaaacga cgatgatca gaagaatta gtttaaagt 
      cattagata aaatctagt ttggtttgt atattttga 
      catgaagtt tctatattt tattgaagt gttgaaacg 
      aaaacgtgt ctttttaca
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY