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Gene information
General information
TC005471 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_007423.3: 12592181..12594159
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
    TC005471
    iBeetle screen iB_00863

    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)

    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: 2 (3 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 abdomen musculature partially not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    Comment: the musculature seems to be ok in the first and in the second abdominal segments.  
    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): less than 30%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 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: 20.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 20.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality early (L5/6) - Number of animals showing the phenotype: 2 (11 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC005471 RA
      MVLTKEYRI CMPITVEEY QVGQLYMIA RHSLEQSGD 
      GEGVEVVEN REIDDPEHG KGQFTEKRI HLSSRLPYW 
      VQAIIPKIF YVTEKAWNY YPYTITEYT CSFIPRFHI 
      EIKTKYENN NGSTENCLN LTPEQLAER LVDHIDIAY 
      DEVSPKHYK EEEDPRFFQ SKKTQRGPL VEGWRANYT 
      PIMCSYKLV NASFEVFGL QTKVEDFIH SCIREVLLL 
      GHRQAFAWI DEWIEMSLQ DVREYEAKL QSQTNSLLA 
      DKVGGGDTA AQESPKSPK SPQKKGYFS WF
    CDS sequence
    • >TC005471 RA
      atggttttg actaaagaa taccgaatt tgtatgccg 
      attacggtc gaagagtat caagtgggg caactgtac 
      atgatcgcg aggcacagt ttggagcag tcgggggat 
      ggcgaagga gttgaagtg gttgaaaat cgggaaatt 
      gacgacccg gagcatggc aaaggccag tttaccgaa 
      aaaagaatt cacttatca agccgtttg ccttactgg 
      gtgcaggcc attatacct aagatattt tacgtgacg 
      gagaaagcg tggaattat tacccgtac actatcaca 
      gaatatacg tgctcattt attccacgt tttcacatt 
      gaaattaaa accaaatat gagaataac aacggcagt 
      accgaaaac tgtcttaat ttaactcct gaacaactt 
      gccgaacgc ctagttgac cacattgac atagcctac 
      gatgaagtg agccccaaa cactacaaa gaggaagag 
      gacccgcgt tttttccag tcgaaaaaa acgcaaagg 
      gggcccctg gtggagggc tggcgggct aactacacc 
      ccaatcatg tgttcgtat aaattagta aatgcctcg 
      tttgaggtt tttggcctc cagacgaaa gtcgaggat 
      tttatccat tcgtgtatt cgcgaggtg ctgttgctg 
      gggcacagg caagccttt gcgtggatt gacgaatgg 
      atagaaatg tcgttgcag gatgtgcgt gaatacgag 
      gcgaagttg caatctcag accaattcg ctgcttgct 
      gataaggtc gggggcggc gatacagcc gcccaggag 
      tcccccaaa agccccaag tcgccccag aaaaagggc 
      tatttttcg tggttctag
    mRNA sequence
    • >TC005471 RA
      agttggcgg gactgcgcg cacgcgcta cattttgtt 
      taaccatac tgtcaaatt tttccaaaa aataaaaac 
      attttttgc gatggtttt gactaaaga ataccgaat 
      ttgtatgcc gattacggt cgaagagta tcaagtggg 
      gcaactgta catgatcgc gaggcacag tttggagca 
      gtcggggga tggcgaagg agttgaagt ggttgaaaa 
      tcgggaaat tgacgaccc ggagcatgg caaaggcca 
      gtttaccga aaaaagaat tcacttatc aagccgttt 
      gccttactg ggtgcaggc cattatacc taagatatt 
      ttacgtgac ggagaaagc gtggaatta ttacccgta 
      cactatcac agaatatac gtgctcatt tattccacg 
      ttttcacat tgaaattaa aaccaaata tgagaataa 
      caacggcag taccgaaaa ctgtcttaa tttaactcc 
      tgaacaact tgccgaacg cctagttga ccacattga 
      catagccta cgatgaagt gagccccaa acactacaa 
      agaggaaga ggacccgcg ttttttcca gtcgaaaaa 
      aacgcaaag ggggcccct ggtggaggg ctggcgggc 
      taactacac cccaatcat gtgttcgta taaattagt 
      aaatgcctc gtttgaggt ttttggcct ccagacgaa 
      agtcgagga ttttatcca ttcgtgtat tcgcgaggt 
      gctgttgct ggggcacag gcaagcctt tgcgtggat 
      tgacgaatg gatagaaat gtcgttgca ggatgtgcg 
      tgaatacga ggcgaagtt gcaatctca gaccaattc 
      gctgcttgc tgataaggt cgggggcgg cgatacagc 
      cgcccagga gtcccccaa aagccccaa gtcgcccca 
      gaaaaaggg ctatttttc gtggttcta gcatcagga 
      ggttacgtt agaggattg ttgcgtttt tgttctttt 
      tttgttact tcatgttta gttgctttt gtttaacta 
      gtcaaatgt ctgcgtcat ttgttatcg cttgacgtt 
      taggaaatt tttatttaa cgaatttta ggtgcactc 
      tggaccagg cgtttcttc tcaattttt tgtatgtat 
      gtagtgtag aagtgagaa tagttgtgt aaaggggat 
      ttttgtctc caacttcct cgtttatcg tgattaata 
      cctacgttg tattgttgg aaaaataaa atttcacac 
      ggaaa
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY