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Gene information
General information
TC002446 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: 29467991..29469578
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
    TC002446
    iBeetle screen iB_00377

    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: 33.33% (includes death as pupa, adult)

    pupa eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (3 dpi)
    Days post injection


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


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


    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 50
    egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - 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): 30% - 50%
    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: 11.11% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)

    pupal molt delayed (still larva) - Number of animals showing the phenotype: 6 (11 dpi)
    Days post injection


    pupal molt delayed (still prepupa) - Number of animals showing the phenotype: 2 (11 dpi)
    Days post injection


    pupal molt not fulfilled (died as L7 or prepupa) - Number of animals showing the phenotype: 6 (19 dpi)
    Days post injection



    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC002446 RA
      MAAPIEALT EAVLQKTKD SSFLKEKVS GYDWSQGLN 
      YDKLLKSYL HTGFQATNF GLAVEEINK MIECRKAPI 
      PEDRYVDNE DPFIGVKNN CAIFLGYTS NIVSSGLRE 
      TIKFLVQHK LVDCIVTTA GGVEEDFIK CMAPTFLGN 
      FALDGVNLR SNGINRIGN LLVPNDNYC QFENWIMPL 
      LDEMLVEQK EGALWSPSK VITRLGERI NNDESIYYW 
      AAKNKIPVF CPALTDGSI GDMMFMHSF RNPGLVLDI 
      ISDLRQLNL MAMKAINTG MIIIGGGVV KHHICNANL 
      MRNGADFCV FLNTASEFD GSDSGARPD EAKSWGKIK 
      KEATPVKIY AEASLIFPL LVAQTFAKE VQIKDK
    CDS sequence
    • >TC002446 RA
      atggcagcc cccattgaa gccctaaca gaggcagtt 
      ttgcaaaaa acaaaagac tcgtcgttt cttaaagaa 
      aaagtgtca ggttacgac tggtcacag ggcttgaac 
      tatgacaaa ctcctcaaa agttatctg cacaccggt 
      tttcaagcc actaatttt ggacttgcc gttgaagaa 
      atcaacaaa atgatcgaa tgtcgcaaa gccccaatt 
      cccgaagac aggtacgtc gataatgag gaccccttt 
      atcggtgtc aagaacaac tgtgcgatt tttttgggg 
      tacacctca aatattgtc tcgagtggc ttacgcgag 
      acaattaag tttctagtg caacacaag ttggttgat 
      tgtattgtt acgacagcg gggggcgta gaggaggac 
      tttatcaag tgtatggcc cccacgttt ctggggaat 
      tttgcgtta gatggggtc aacttaagg agtaatggg 
      atcaataga atcgggaat ttgttagtg cctaatgat 
      aattattgc caattcgag aactggatt atgcccctt 
      ctggacgaa atgttagtt gaacagaaa gagggggct 
      ttgtggagc cccagtaaa gtcataact aggctaggt 
      gaaaggata aataacgac gaaagtatt tattactgg 
      gctgctaaa aataaaatc cctgttttt tgcccggct 
      ttgactgat ggaagcata ggcgatatg atgtttatg 
      cactcattc cgaaatccg ggcctggtt ttggacatt 
      atttcggac ttgagacag ttaaattta atggcaatg 
      aaggcaatc aacaccggg atgattata atcgggggt 
      ggcgtggtc aagcaccac atttgcaat gccaattta 
      atgcgaaat ggtgccgat ttttgcgtg tttctgaac 
      acagcctcc gagtttgac ggcagtgac tcgggggcc 
      cgccccgac gaggccaaa tcctggggc aaaataaaa 
      aaagaagcc actcctgtg aaaatctac gccgaagcc 
      agcttgatt ttcccactt ttggtggcc caaactttc 
      gcaaaggag gtccaaatc aaagacaaa tag
    mRNA sequence
    • >TC002446 RA
      tggggttag gtgtgggca tggcagccc ccattgaag 
      ccctaacag aggcagttt tgcaaaaaa caaaagact 
      cgtcgtttc ttaaagaaa aagtgtcag gttacgact 
      ggtcacagg gcttgaact atgacaaac tcctcaaaa 
      gttatctgc acaccggtt ttcaagcca ctaattttg 
      gacttgccg ttgaagaaa tcaacaaaa tgatcgaat 
      gtcgcaaag ccccaattc ccgaagaca ggtacgtcg 
      ataatgagg accccttta tcggtgtca agaacaact 
      gtgcgattt ttttggggt acacctcaa atattgtct 
      cgagtggct tacgcgaga caattaagt ttctagtgc 
      aacacaagt tggttgatt gtattgtta cgacagcgg 
      ggggcgtag aggaggact ttatcaagt gtatggccc 
      ccacgtttc tggggaatt ttgcgttag atggggtca 
      acttaagga gtaatggga tcaatagaa tcgggaatt 
      tgttagtgc ctaatgata attattgcc aattcgaga 
      actggatta tgccccttc tggacgaaa tgttagttg 
      aacagaaag agggggctt tgtggagcc ccagtaaag 
      tcataacta ggctaggtg aaaggataa ataacgacg 
      aaagtattt attactggg ctgctaaaa ataaaatcc 
      ctgtttttt gcccggctt tgactgatg gaagcatag 
      gcgatatga tgtttatgc actcattcc gaaatccgg 
      gcctggttt tggacatta tttcggact tgagacagt 
      taaatttaa tggcaatga aggcaatca acaccggga 
      tgattataa tcgggggtg gcgtggtca agcaccaca 
      tttgcaatg ccaatttaa tgcgaaatg gtgccgatt 
      tttgcgtgt ttctgaaca cagcctccg agtttgacg 
      gcagtgact cgggggccc gccccgacg aggccaaat 
      cctggggca aaataaaaa aagaagcca ctcctgtga 
      aaatctacg ccgaagcca gcttgattt tcccacttt 
      tggtggccc aaactttcg caaaggagg tccaaatca 
      aagacaaat agaaacttt ataatttta ttaataaaa 
      taatttcac tgct
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY