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Gene information
General information
TC003566 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: 7605359..7606505
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
    TC003566
    iBeetle screen iB_10960

    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)

    adult lethality - Number of animals showing the phenotype: 1 (9 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: < 50 embryo/egg with embryonic tissue potentially present, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): less than 30%

    embryo/egg not developed/not fertilized - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    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: > 50
    embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    cuticle remnants with some identifiable segments - Penetrance (percentage of animals/eggs on slide): less than 30%

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

    head shape irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    head not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    thorax shape irregular - Penetrance (percentage of animals/eggs on slide): less than 30%
    thorax not present - Penetrance (percentage of animals/eggs on slide): less than 30%
    abdomen shape irregular - Penetrance (percentage of animals/eggs on slide): less than 30%
    abdomen not present - Penetrance (percentage of animals/eggs on slide): less than 30%
    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 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
    • >TC003566 RA
      MQDPNEDTE WNDVLRSKG IIPPKKETE 
      ITEDQIVSM LEETIEQKT STGKDFSKL 
      DLDELDELE DSEDEAVLL EYRNKRIAE 
      MRALAEKAK FGSVGEISA QDYVQEVNK 
      AGTGIWVVL HLYKQGIPL CALINEYMK 
      QLAAKYPAV KFLKSISTT CIPNYPDKN 
      LPTLFVYFE GELKSQIVG PVELRGPNM 
      TQDEFEFLI GKTGAIETS IKEDPKPKI 
      KDKMFADLA STNDW
    CDS sequence
    • >TC003566 RA
      atgcaggac cccaatgag gacactgaa tggaacgat 
      gttctcaga agcaaggga attattcct cctaagaag 
      gaaacagag ataacagag gatcaaatt gtgtctatg 
      ttggaagag acaattgaa caaaaaacc tcaactggt 
      aaagatttt tcaaagcta gaccttgat gagctagac 
      gagttggaa gattctgaa gatgaagct gttcttctt 
      gaatacagg aataagagg atagctgaa atgagagca 
      ttggctgaa aaagccaaa tttggctca gtaggtgaa 
      atttcagcc caagattat gtgcaggaa gttaataaa 
      gctggcaca ggcatttgg gttgtttta catttatac 
      aaacaagga ataccgctg tgtgcccta attaatgaa 
      tatatgaag caacttgca gctaaatac ccagctgtt 
      aagttcctg aaatcaatt tcaacgact tgtatacca 
      aattatccc gacaaaaac ttaccaact ctctttgtg 
      tactttgaa ggggaatta aaatcgcaa attgtgggt 
      ccagtcgaa cttaggggc ccaaatatg acccaagac 
      gagtttgaa tttttaatt ggaaaaaca ggcgctatt 
      gagaccagt attaaagaa gacccaaag ccgaaaatc 
      aaagataaa atgttcgca gatttggct agtaccaat 
      gattggtaa
    mRNA sequence
    • >TC003566 RA
      agatgacaa tatgttacg tgacggcaa aacatcgga 
      tgtgtcatg cctactaca caaatttcc ttttttgta 
      aagtagtca catgtagag cttcaacag taaataatt 
      caaaatttt aatattttt gactcatac agtttaaca 
      aaatgcagg accccaatg aggacactg aatggaacg 
      atgttctca gaagcaagg gaattattc ctcctaaga 
      aggaaacag agataacag aggatcaaa ttgtgtcta 
      tgttggaag agacaattg aacaaaaaa cctcaactg 
      gtaaagatt tttcaaagc tagaccttg atgagctag 
      acgagttgg aagattctg aagatgaag ctgttcttc 
      ttgaataca ggaataaga ggatagctg aaatgagag 
      cattggctg aaaaagcca aatttggct cagtaggtg 
      aaatttcag cccaagatt atgtgcagg aagttaata 
      aagctggca caggcattt gggttgttt tacatttat 
      acaaacaag gaataccgc tgtgtgccc taattaatg 
      aatatatga agcaacttg cagctaaat acccagctg 
      ttaagttcc tgaaatcaa tttcaacga cttgtatac 
      caaattatc ccgacaaaa acttaccaa ctctctttg 
      tgtactttg aaggggaat taaaatcgc aaattgtgg 
      gtccagtcg aacttaggg gcccaaata tgacccaag 
      acgagtttg aatttttaa ttggaaaaa caggcgcta 
      ttgagacca gtattaaag aagacccaa agccgaaaa 
      tcaaagata aaatgttcg cagatttgg ctagtacca 
      atgattggt aatgcggca ataatcatc tttatctac 
      ttaacgaat tttgtaaca attttatta tttattgct 
      gttgctgta ttgaataaa ttagttttt tgatc
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY