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Gene information
General information
TC003775 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: 17770835..17771609
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
    TC003775
    iBeetle screen iB_03619

    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: 0.0% (includes death as pupa, adult)

    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 abdomen musculature lateral orientation irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    Comment: oblique lateral muscles not parellel to each other
    abdomen musculature lateral sometimesnot present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    Comment: one of the two oblique muscles sometimes in single abdominal segments misssing
    abdomen musculature dorsal potentiallythinner - Penetrance (percentage of animals/eggs on slide): less than 30%
    abdomen musculature ventral potentiallythinner - 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
    legs pretarsus orientation irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    legs tibiotarsus shortened - 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: 20.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)

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


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


    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 6 (16 dpi)
    Days post injection



    Pupal morphology
    antenna shape rounded - Number of animals showing the phenotype: 8

    elytra orientation irregular - Number of animals showing the phenotype: 2

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC003775 RA
      MSITIEKTK YECAALPKR WQREDMLRK 
      TGISVGKVD VMYYSRGVR NDASLVPPI 
      RQTASIFKQ PVTVYKTQE SKVKSDYKN 
      GNQEKPKQL IWEKRLEGL RACDLDGTE 
      FEAMELPKG LKPVGPNVS EETIIQSVA 
      TALHVSTQP VTGQTGPKA ALEKNPGVF 
      LDPKQPLVQ AVNISDEDI KRQEDRVAF 
      ARKKLQEAL KS
    CDS sequence
    • >TC003775 RA
      atgagtata actatcgaa aaaacgaaa tatgagtgt 
      gccgcgttg cccaagcga tggcagcgc gaggatatg 
      ttgcgcaaa acggggatt tccgtcggc aaagtggac 
      gtgatgtat tacagtcgg ggcgtccgc aatgacgct 
      tccttggtg ccccccatc cggcagacg gcgtcgatt 
      ttcaagcag ccggtcact gtgtacaag acacaggag 
      agtaaggtg aagtccgac tacaaaaac gggaatcag 
      gagaagccg aaacagttg atttgggag aagaggttg 
      gaggggttg cgggcgtgc gatttggac gggacggag 
      ttcgaggcg atggagttg cccaagggg ctgaagccg 
      gtggggccc aatgtgagt gaggagacg attatacag 
      agtgtggcc acagcgttg catgtctcc acgcaaccc 
      gtgacgggg caaacgggg cccaaagcc gccttggag 
      aagaatccg ggggtgttt ttggacccc aaacagccc 
      ttggtccaa gcggtgaat atctccgat gaggatatt 
      aaaaggcag gaggatagg gtggctttt gccaggaaa 
      aagttgcaa gaggcttta aagtcgtag
    mRNA sequence
    • >TC003775 RA
      gttggcgtc ccgcatcag ctgtgtttt gtgtacggt 
      gtcaataaa tttgacaat tttgccaaa aatcgctca 
      aaaaatgag tataactat cgaaaaaac gaaatatga 
      gtgtgccgc gttgcccaa gcgatggca gcgcgagga 
      tatgttgcg caaaacggg gatttccgt cggcaaagt 
      ggacgtgat gtattacag tcggggcgt ccgcaatga 
      cgcttcctt ggtgccccc catccggca gacggcgtc 
      gattttcaa gcagccggt cactgtgta caagacaca 
      ggagagtaa ggtgaagtc cgactacaa aaacgggaa 
      tcaggagaa gccgaaaca gttgatttg ggagaagag 
      gttggaggg gttgcgggc gtgcgattt ggacgggac 
      ggagttcga ggcgatgga gttgcccaa ggggctgaa 
      gccggtggg gcccaatgt gagtgagga gacgattat 
      acagagtgt ggccacagc gttgcatgt ctccacgca 
      acccgtgac ggggcaaac ggggcccaa agccgcctt 
      ggagaagaa tccgggggt gtttttgga ccccaaaca 
      gcccttggt ccaagcggt gaatatctc cgatgagga 
      tattaaaag gcaggagga tagggtggc ttttgccag 
      gaaaaagtt gcaagaggc tttaaagtc gtagagtgt 
      agatgaatt ttatttatt ttaggtgta tcatgtgac 
      taggtttaa gttttgtta agcgatttt tttaataca 
      ataaagttg attaaggat a
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY