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Gene information
General information
TC030221 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: 16042618..16045691
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
    TC030221
    iBeetle screen iB_07454

    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: 70.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 5 (3 dpi)
    Days post injection


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


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


    adult nutritional state potentially bad (cachectic) - Number of animals showing the phenotype: 2 (11 dpi)
    Days post injection


    adult nutritional state bad (cachectic) - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection


    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
    • >TC030221 RA
      MNQFVEPVT LLIFYYIVP ITIKCFDNL FKRDILNLL 
      LVKFKFLNK EFDRQSDIA DLPKLFPLT KTKKIKFLE 
      NEEIDSNLR YIEELSQYH YHLVQLSLD ICKIFEIEV 
      VITLILWFE KIIESTYFA IFTLVNDDY NHKTLFHAL 
      DVIHATFEF YWLFTSIEN FNRVQIEAN KTPILAHDV 
      WNKYVTKKF DDKVRHLQL ISVRFLNTK LQFTAAGLF 
      NLDRSFCHT VIFGVTVYV VILIQFYIY I
    CDS sequence
    • >TC030221 RA
      atgaatcag tttgtagaa ccagtcact cttttaatt 
      ttttactac attgttcct attacgata aagtgtttt 
      gacaacttg ttcaaaaga gatatacta aacctactt 
      ttagttaaa tttaaattt cttaacaaa gaatttgat 
      cgacaatcc gatattgct gatttacca aaattgttt 
      cctttgact aaaaccaaa aaaattaag tttttggaa 
      aatgaagaa attgattcg aatctcaga tatattgaa 
      gaactctca caatatcac taccatcta gtccagtta 
      tcactcgat atttgcaaa atttttgaa attgaagtt 
      gtgatcact ctaatactg tggtttgag aagattatt 
      gaaagcacc tattttgct atatttact ttagtaaac 
      gatgactat aatcataaa acgctgttt catgctctt 
      gatgttatt catgcaact tttgaattc tattggctc 
      tttacatcc atcgagaat ttcaatcgc gtccaaatt 
      gaagccaat aaaacaccg attcttgca cacgacgtt 
      tggaacaag tatgttacg aaaaaattt gacgacaaa 
      gtgcgacat ctacaatta atttcagtc cgatttcta 
      aacacaaaa ttacaattt acagcagcc ggattgttt 
      aatttagac cggagtttc tgccataca gtaattttt 
      ggagtaacc gtttacgtc gtgatttta attcagttt 
      tatatttac atctga
    mRNA sequence
    • >TC030221 RA
      atcttttga atgaatcag tttgtagaa ccagtcact 
      cttttaatt ttttactac attgttcct attacgata 
      aagtgtttt gacaacttg ttcaaaaga gatatacta 
      aacctactt ttagttaaa tttaaattt cttaacaaa 
      gaatttgat cgacaatcc gatattgct gatttacca 
      aaattgttt cctttgact aaaaccaaa aaaattaag 
      tttttggaa aatgaagaa attgattcg aatctcaga 
      tatattgaa gaactctca caatatcac taccatcta 
      gtccagtta tcactcgat atttgcaaa atttttgaa 
      attgaagtt gtgatcact ctaatactg tggtttgag 
      aagattatt gaaagcacc tattttgct atatttact 
      ttagtaaac gatgactat aatcataaa acgctgttt 
      catgctctt gatgttatt catgcaact tttgaattc 
      tattggctc tttacatcc atcgagaat ttcaatcgc 
      gtccaaatt gaagccaat aaaacaccg attcttgca 
      cacgacgtt tggaacaag tatgttacg aaaaaattt 
      gacgacaaa gtgcgacat ctacaatta atttcagtc 
      cgatttcta aacacaaaa ttacaattt acagcagcc 
      ggattgttt aatttagac cggagtttc tgccataca 
      gtaattttt ggagtaacc gtttacgtc gtgatttta 
      attcagttt tatatttac atctgaaaa taaaatgct 
      caaacaaaa
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY