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Gene information
General information
TC004961 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: 5269866..5279072
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
    TC004961
    iBeetle screen iB_03883

    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


    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: 10.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 30.0% (includes death as larva, prepupa, pupa, adult)

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


    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 2 (16 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC004961 RA
      MLETPVMSE SAFRARDIG LRAQKKILS 
      RMAGKNVAR AFIDDTTAS LLDNLYRLA 
      KQYSQNKKE AEKLIKNII KVVIKLGVL 
      HRNGQFSSE ELKQAERFK AKFRMAGMA 
      IISFYEVDF SYDRNYLTA ALAESHKCL 
      QFIVSKHLT EKSLSRIDS VFSFFSNEQ 
      FLDDIFQVN SEYRETLGR IVADLNKAI 
      ENGDL
    CDS sequence
    • >TC004961 RA
      atgttggaa acaccagtc atgagcgag agcgcattt 
      agggcgcgt gacatcggc cttcgagcc cagaaaaag 
      attttgtct cgcatggcc ggtaaaaac gtggctcgt 
      gcattcatc gacgacacc actgcgtcc ctattggat 
      aacttatac cgtcttgcc aagcaatac tcccagaat 
      aagaaggaa gcggaaaag ctgataaag aacatcatc 
      aaagtggtg attaaactt ggggttttg caccggaac 
      ggccagttt tcaagcgag gaattgaaa caagcggag 
      cgcttcaaa gcaaagttt cgcatggct ggaatggcg 
      atcatatca ttctacgaa gtggacttc agctatgat 
      aggaattat ctgactgcg gctcttgca gagtcacac 
      aagtgcctc cagtttatc gtgtccaag catcttacc 
      gaaaaatcc ttatcgcgc atcgacagc gtcttcagt 
      ttcttctca aatgaacag ttcttagat gatattttt 
      caagtgaat tcggagtac agagaaacg ttggggaga 
      attgtcgca gacctcaat aaagccatc gaaaatggc 
      gatctttag
    mRNA sequence
    • >TC004961 RA
      gagtagcga gtggcagtg agtgtgttg taaacaatc 
      atttctttt gaaattcat aaataagcg aaattttat 
      tgatttgta ttgacatca gcaggacgc actgtaaca 
      ttcccgcta atcggctct tttaatttt atgctctct 
      acacgtctt tcactgccc tttaagttg cctccgaaa 
      tgacgacga tggtggaac tcgaaagtt ggctcattt 
      atcagtgca ttgcgacag aatgcagca actgcttgt 
      gtaagccac aatcacaac aaccaccac cgtttaact 
      tatttttga gttgcttta tttttcttt cattttttt 
      aggattttt tgagttggc cctattttt aaacactag 
      gtctaatgt tggaaacac cagtcatga gcgagagcg 
      catttaggg cgcgtgaca tcggccttc gagcccaga 
      aaaagattt tgtctcgca tggccggta aaaacgtgg 
      ctcgtgcat tcatcgacg acaccactg cgtccctat 
      tggataact tataccgtc ttgccaagc aatactccc 
      agaataaga aggaagcgg aaaagctga taaagaaca 
      tcatcaaag tggtgatta aacttgggg ttttgcacc 
      ggaacggcc agttttcaa gcgaggaat tgaaacaag 
      cggagcgct tcaaagcaa agtttcgca tggctggaa 
      tggcgatca tatcattct acgaagtgg acttcagct 
      atgatagga attatctga ctgcggctc ttgcagagt 
      cacacaagt gcctccagt ttatcgtgt ccaagcatc 
      ttaccgaaa aatccttat cgcgcatcg acagcgtct 
      tcagtttct tctcaaatg aacagttct tagatgata 
      tttttcaag tgaattcgg agtacagag aaacgttgg 
      ggagaattg tcgcagacc tcaataaag ccatcgaaa 
      atggcgatc tttagtgaa gcttcagcg acggttttt 
      gtgattttg ttgtgattt taatttagt tgttttgtg 
      tattattgg acgtcgtag ttgttgcat gggactagg 
      tttgtgtta tattgtacc taagaatct atttttatc 
      gttgccttc tttacaagt ttcgttcga cgttcagtg 
      tgttgataa atgttaccc tttcagaga gtttttcgg 
      gagcgttag gagtggtcc aatcaatct tatttaacg 
      taattaagc aacagaatt ttgaatcga acatcatat 
      ctatactaa aagtaacca aacattttt accaaaatc 
      tatttttat atgaccaaa attctgttg ctctctgtt 
      ttgtcttct ctaaagaaa cgcttaaaa agacacatg 
      cttaaccac actatgtag atcgtagtt tctggtcca 
      cgtttcatg atttttgta aagacataa ggcacgtgt 
      acatattaa ttgacatat cttttattt acacgattt 
      aacgatatt gtagactaa gatattttt gcaccaaga 
      aatttaaaa ggccggtca aatcactat tcacgagcc 
      taaaactgt tgtgcattt ctaataaaa gtaaaaaca 
      gtaaagaca ataagtcag tcatcttct attcatttt 
      ctactagtc ataagtcgt ttatttcct atttattga 
      gttaagcca aatattgaa ttaagtttt attgttttt 
      atgacaggg cgatttttg tgaccgagt tgttcttgc 
      ctggttttt caaacttga atgaatatc agtgatgaa 
      actcatgtt tgcatgaaa atataattc caattattg 
      ttacactat aaggaatag tagatctca gatattcgg 
      caatgtatg atgttttca atgtttttc tgtatttag 
      ttgttcgtt atcaaaata ataaacaac atctaatac 
      g
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