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Gene information
General information
TC014559 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: 11772851..11773745
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
    TC014559
    iBeetle screen iB_02302

    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: 50.0% (includes death as pupa, adult)

    pupa eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 3 (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


    Adult morphology
    wings not closed - Number of animals showing the phenotype: 3 (13 dpi)
    Days post injection


    elytra blistered - Number of animals showing the phenotype: 3 (13 dpi)
    Days post injection


    hindwing irregular - Number of animals showing the phenotype: 3 (13 dpi)
    Days post injection


    elytra surface irregular - Number of animals showing the phenotype: 3 (13 dpi)
    Days post injection



    Fertility
    mature egg deposition blocked number 1st lateral oviduct: 2 number 2nd lateral oviduct: >5 - Number of animals showing the phenotype: 1
    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: 40.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 44.44% (includes death as larva, prepupa, pupa, adult)

    larva lethality early (L5/6) - Number of animals showing the phenotype: 4 (11 dpi)
    Days post injection


    Pupal morphology
    head bristle mostly decreased - Number of animals showing the phenotype: 5

    abdomen bristle partially not present - Number of animals showing the phenotype: 5

    Adult morphology
    thorax sticky - Number of animals showing the phenotype: 4


    abdomen sticky - Number of animals showing the phenotype: 4

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC014559 RA
      MSASTSPSH YTEDAFVTD ANFKVEINQ 
      KMKVPDKIS FNTDLNGAT QPPWNRDNI 
      NMQVPERIL VVGQHQHIG TRAPPREIV 
      FDNSILPPE PYPGDVRVA TPPRTLTLD 
      KYPFPTLEE MEDPELNQI PVVKPKKKQ 
      AFDDSDLNL TIRETTPPI GGSGERLTA 
      AEEVLHLRR QMAKLNRRV MALELENLS 
      RLQKEKFLV GFGVAYLLL KVIIWMNRD
    CDS sequence
    • >TC014559 RA
      atgtcagcg agcacaagt ccgtcgcat tacaccgaa 
      gacgccttc gtgacagat gccaacttc aaagtggaa 
      attaatcaa aaaatgaaa gttcccgac aagatcagc 
      ttcaacacg gacttgaac ggggccaca cagcccccg 
      tggaaccgc gacaacatc aacatgcaa gtccctgag 
      cgaatcctc gtcgtgggg caacatcag cacattggg 
      acaagggcg cccccacgc gaaattgtc tttgacaac 
      tcgatcctt cccccagaa ccgtacccc ggggatgtg 
      cgggttgcg accccgccc aggactttg actctggac 
      aagtacccc tttcccact cttgaggag atggaggac 
      ccggaactg aaccagatt ccggtggtt aaaccaaag 
      aaaaaacag gcgtttgat gattcggat ttgaatctg 
      acgatacgc gagactacg cccccgatc gggggcagc 
      ggcgagagg ctgacggcc gccgaagag gtgctgcac 
      ctcaggcgg caaatggcg aagctcaac agacgtgtg 
      atggcctta gaattagaa aacttgagc cggctccaa 
      aaggagaag tttctagtt ggttttgga gtcgcttat 
      ttgttgttg aaagtcatt atttggatg aatagagat 
      tag
    mRNA sequence
    • >TC014559 RA
      tcgctgata aaaatagca ttatttggc tataaatag 
      cgttatcac accagtaaa agatgtcag cgagcacaa 
      gtccgtcgc attacaccg aagacgcct tcgtgacag 
      atgccaact tcaaagtgg aaattaatc aaaaaatga 
      aagttcccg acaagatca gcttcaaca cggacttga 
      acggggcca cacagcccc cgtggaacc gcgacaaca 
      tcaacatgc aagtccctg agcgaatcc tcgtcgtgg 
      ggcaacatc agcacattg ggacaaggg cgcccccac 
      gcgaaattg tctttgaca actcgatcc ttcccccag 
      aaccgtacc ccggggatg tgcgggttg cgaccccgc 
      ccaggactt tgactctgg acaagtacc cctttccca 
      ctcttgagg agatggagg acccggaac tgaaccaga 
      ttccggtgg ttaaaccaa agaaaaaac aggcgtttg 
      atgattcgg atttgaatc tgacgatac gcgagacta 
      cgcccccga tcgggggca gcggcgaga ggctgacgg 
      ccgccgaag aggtgctgc acctcaggc ggcaaatgg 
      cgaagctca acagacgtg tgatggcct tagaattag 
      aaaacttga gccggctcc aaaaggaga agtttctag 
      ttggttttg gagtcgctt atttgttgt tgaaagtca 
      ttatttgga tgaatagag attagggag cactagttt 
      atccctaag ttaggaaca aatcgtatt gacgttaat 
      tagttctgg tttaagtaa cataattt
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY