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Gene information
General information
TC009363 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_007422.5: 1057936..1059134
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
    TC009363
    iBeetle screen iB_08599

    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: 20.0% (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


    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): 50% - 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    remarks: muscle pattern potentially obscured by segmentation defect,
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    larva partially everted (inside-out) - 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
    • >TC009363 RA
      MKVCICFAL LLSCAHSSD FQKWENFKK KFNKTYDTK 
      LEENHRRTI FFSNLQLIE NHNEKYENE SKFVPKTKV 
      PESIDWRDY GVITPIVDQ NYCLSCWAF AVVAALEGH 
      VGIHLGQKN QTLSQENLI DCVYSDFQC KQEMKRSAL 
      VDCYQYMVN SGGIDTLES YPYDQKPPL CRFKPENIG 
      ASIQGYGTV TEGDEEELK AVVGTLGPV SVIVTADLI 
      FILYRKGIY FNDNWLNAS EPYNHALTV IGYGSENGQ 
      DYWIVRNE
    CDS sequence
    • >TC009363 RA
      atgaaagtt tgtatttgc ttcgcttta ttgctcagc 
      tgcgcccat tcgagtgat ttccaaaaa tgggaaaac 
      tttaaaaaa aaattcaac aaaacttat gacaccaaa 
      ttggaagag aaccaccga cgaacaatt tttttcagc 
      aacctccaa ctgattgaa aatcacaat gaaaaatat 
      gagaacgag agtaaattt gtaccaaaa accaaagtc 
      cccgaaagt atcgactgg agggattat ggtgtcatt 
      acaccaatt gtggaccaa aactattgc ttgtcgtgt 
      tgggctttt gctgtggtg gcggcatta gaaggccac 
      gttggcata caccttggt caaaaaaac caaactctg 
      agtcaggaa aacttgatt gattgcgtc tactccgat 
      ttccagtgc aagcaagaa atgaaaagg tcagctctg 
      gttgattgt taccaatac atggtcaac agcggaggg 
      atcgatact ttggaatca tacccttac gaccagaaa 
      ccacccctg tgtcgtttc aagccggaa aatattggt 
      gccagtatc caaggatat ggtactgtt actgaaggt 
      gatgaggaa gagttgaag gcagttgtg gggacactt 
      ggacccgtt tctgtgatt gtaacagct gatcttatc 
      tttatcttg taccgtaag ggaatttac tttaatgac 
      aattggttg aacgcctca gaaccctac aatcatgca 
      cttactgta attggttac ggttccgaa aacgggcaa 
      gattattgg attgtgcga aacgaatga
    mRNA sequence
    • >TC009363 RA
      agataaatt tatccgaaa agaagcagt tgacaacag 
      gtttatgca ctcattaaa ctgtatttc ggcaatgaa 
      agtttgtat ttgcttcgc tttattgct cagctgcgc 
      ccattcgag tgatttcca aaaatggga aaactttaa 
      aaaaaaatt caacaaaac ttatgacac caaattgga 
      agagaacca ccgacgaac aattttttt cagcaacct 
      ccaactgat tgaaaatca caatgaaaa atatgagaa 
      cgagagtaa atttgtacc aaaaaccaa agtccccga 
      aagtatcga ctggaggga ttatggtgt cattacacc 
      aattgtgga ccaaaacta ttgcttgtc gtgttgggc 
      ttttgctgt ggtggcggc attagaagg ccacgttgg 
      catacacct tggtcaaaa aaaccaaac tctgagtca 
      ggaaaactt gattgattg cgtctactc cgatttcca 
      gtgcaagca agaaatgaa aaggtcagc tctggttga 
      ttgttacca atacatggt caacagcgg agggatcga 
      tactttgga atcataccc ttacgacca gaaaccacc 
      cctgtgtcg tttcaagcc ggaaaatat tggtgccag 
      tatccaagg atatggtac tgttactga aggtgatga 
      ggaagagtt gaaggcagt tgtggggac acttggacc 
      cgtttctgt gattgtaac agctgatct tatctttat 
      cttgtaccg taagggaat ttactttaa tgacaattg 
      gttgaacgc ctcagaacc ctacaatca tgcacttac 
      tgtaattgg ttacggttc cgaaaacgg gcaagatta 
      ttggattgt gcgaaacga atgaagccg agtatttgg 
      agaagatgg ttacatcag aatggccag aaatcggga 
      caacaattg tggaattgc aacttacgc cagttatcc 
      cgttgtaaa taaataatt aggtttttg tagttgtaa 
      taataataa aatcttgat gggtca
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY