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Gene information
General information
TC014474 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_007420.3: 10824178..10825573
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
    TC014474
    iBeetle screen iB_05637

    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: 30.0% (includes death as pupa, adult)

    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: 2 (3 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 embryo/egg with muscle fluorescence/pattern, but not hatched (pot. late embryonic lethality) - Penetrance (percentage of animals/eggs on slide): less than 30%
    larval musculature ventral partiallynot present - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    larval musculature lateral partiallynot present - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    larva segment ventral potentiallyfused - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - 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: 10.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 37.5% (includes death as larva, prepupa, pupa, adult)

    adult lethality - Number of animals showing the phenotype: 2 (22 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 1 (41 dpi)
    Days post injection


    Adult morphology
    ommatidia partially pigmentation/tanning not present - Number of animals showing the phenotype: 8

    Comment: outer ring of ommatidia lacks black pigmentation

    eyes potentially slightlysize decreased - Number of animals showing the phenotype: 8

    Fertility
    egg production: number <50% of expected -
    mature egg deposition 2 number 2nd lateral oviduct: 5 number 1st lateral oviduct: blocked - Number of animals showing the phenotype: 3
    mature egg deposition 2 number 2nd lateral oviduct: 3 number 1st lateral oviduct: blocked - Number of animals showing the phenotype: 1
    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC014474 RA
      MLDMPGTIM AKNYSHCPL KKRPVQMLE DIPDEPENL 
      STKPEDLSR TGRLASVSP SSLSPVSPN SSVKTSPPP 
      SPPMRSPSP HYYPPKTEI LPYQEYAPQ TWHRSVAPL 
      YPPVYHPFV DYMGMSPTY SESSLSPPH HHVQQPLQP 
      LALRPYQLD QASLSPSSW TTSPPPNTT PEDLSEKRR 
      QGVRHQCPD CGKSYSTFS GLSKHRQFH CAAGEGPKK 
      SFSCKFCEK VYVSLGALK MHIRTHTLP CKCTICGKA 
      FSRPWLLQG HIRTHTGEK PFSCTYCNR AFADRSNLR 
      AHLQTHSDV KKYSCPTCS KTFSRMSLL TKHAEGGCS 
      GVHQEKMY
    CDS sequence
    • >TC014474 RA
      atgttggat atgcccgga accatcatg gccaaaaac 
      tactcgcat tgcccgctc aagaaacga cccgtgcaa 
      atgctggag gacatccct gacgaaccg gaaaatctc 
      agcacgaaa ccggaagat ttgtcccgg accggccgt 
      ctggcctca gtctcgcct tcctcccta tctccggtc 
      tccccgaac tcgtcagtg aaaacctcg ccgccccca 
      tcgccccca atgcgctcg ccgtcccct cactactac 
      cccccgaaa accgagatc ttgccatac caggaatac 
      gccccccag acctggcac cgcagcgtc gcccctttg 
      taccctcca gtgtaccac cccttcgtg gactacatg 
      gggatgagc cccacgtac agcgaaagc tcgctctcg 
      ccgccgcac caccacgtc cagcagccc ctgcagccc 
      ctagccttg cgcccttat cagctcgac caagcctcc 
      ctgagcccc agttcttgg acgacttcg ccgcccccc 
      aacaccacc cccgaggac ctgtcggag aagcgccgc 
      cagggtgtg cgccaccag tgccccgac tgtggcaaa 
      agctactcc actttctcc gggctttcc aagcaccgt 
      cagttccac tgcgctgcc ggtgaaggc ccgaagaag 
      tccttcagt tgcaaattc tgtgagaaa gtttacgtt 
      tctttgggc gccctcaag atgcacatc aggacccac 
      acgttgccc tgcaagtgc accatctgc gggaaggcc 
      ttctccagg ccgtggctg ctccaaggg cacattagg 
      acgcacacg ggcgagaaa cccttctct tgcacttat 
      tgtaatcgg gcgtttgcc gaccggtcc aacttgaga 
      gcgcatctt cagacacat tccgatgtg aagaagtat 
      tcgtgtccc acctgcagt aaaaccttc tcgaggatg 
      agtctgctg acgaagcat gcggaaggc ggctgcagc 
      ggcgtgcac caggagaag atgtactag
    mRNA sequence
    • >TC014474 RA
      actcgctcg aatgttcat gtgttgtgc cagtgactc 
      gcttggatt attgtgaag tgatgttgg atatgcccg 
      gaaccatca tggccaaaa actactcgc attgcccgc 
      tcaagaaac gacccgtgc aaatgctgg aggacatcc 
      ctgacgaac cggaaaatc tcagcacga aaccggaag 
      atttgtccc ggaccggcc gtctggcct cagtctcgc 
      cttcctccc tatctccgg tctccccga actcgtcag 
      tgaaaacct cgccgcccc catcgcccc caatgcgct 
      cgccgtccc ctcactact accccccga aaaccgaga 
      tcttgccat accaggaat acgcccccc agacctggc 
      accgcagcg tcgcccctt tgtaccctc cagtgtacc 
      accccttcg tggactaca tggggatga gccccacgt 
      acagcgaaa gctcgctct cgccgccgc accaccacg 
      tccagcagc ccctgcagc ccctagcct tgcgccctt 
      atcagctcg accaagcct ccctgagcc ccagttctt 
      ggacgactt cgccgcccc ccaacacca cccccgagg 
      acctgtcgg agaagcgcc gccagggtg tgcgccacc 
      agtgccccg actgtggca aaagctact ccactttct 
      ccgggcttt ccaagcacc gtcagttcc actgcgctg 
      ccggtgaag gcccgaaga agtccttca gttgcaaat 
      tctgtgaga aagtttacg tttctttgg gcgccctca 
      agatgcaca tcaggaccc acacgttgc cctgcaagt 
      gcaccatct gcgggaagg ccttctcca ggccgtggc 
      tgctccaag ggcacatta ggacgcaca cgggcgaga 
      aacccttct cttgcactt attgtaatc gggcgtttg 
      ccgaccggt ccaacttga gagcgcatc ttcagacac 
      attccgatg tgaagaagt attcgtgtc ccacctgca 
      gtaaaacct tctcgagga tgagtctgc tgacgaagc 
      atgcggaag gcggctgca gcggcgtgc accaggaga 
      agatgtact agttctgtc aggacttga ggttccttt 
      ttttgacag tattttcgt aaatagtgc ctttttcga 
      agactgaca gtttggaaa tgttatgta caaagtaat 
      tcctaccat gtatatagt aagttaaga aatttaata 
      tatttttgt aaatacaca gacggctcg gatgagtcg 
      cgttctata ctctagtca atacgttat tcttatgcc 
      attgtatag tgccttact tagtcttat tgaacctat 
      attgcagat tttatacta ttatttaat aaaacacat 
      ccacaaaa
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY