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Gene information
General information
TC032499 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: 14087710..14094507
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
    TC032499
    iBeetle screen iB_08010

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

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC032499 RA
      MDFLIGGLA AMGATFFSN PLDVLKTRI QLQGELKPR 
      GQHEIHYRH ALHAAYVVI KQEGIFALQ KGLGAALLM 
      HGVRNSIKL GSYQWFTNQ GYICDSDNR TIFHKSLLA 
      SAFGGAAGA FVGSPLYQI TTQLQSNAG EGIAVGHQH 
      KHSGLVQAF KAIYSRHGM KGLWRGSNA NVIRTVVGA 
      SAQLTTFSK TKDILREYE FFQRSTTMT AFFASIAGG 
      ICQTVFQTP LDLVCIRLN NQYVYASGQ GALYTGMTN 
      CLTKIIQSE GFFGLYKGA GVNYMRIAR QTGFCLMFW 
      ELLKDLHVK HFGLNKNGH CNTSISDEI IKL
    CDS sequence
    • >TC032499 RA
      atggacttc ctaattggc ggtttagct gcaatggga 
      gccactttt ttcagcaat ccccttgac gttcttaaa 
      actagaata cagcttcaa ggagaactg aaacctcga 
      gggcaacat gaaatccac tacagacac gcactgcat 
      gctgcttat gttgtaata aaacaggaa ggaattttt 
      gcattgcag aaaggactt ggagctgca cttctgatg 
      catggagtg cgtaacagt atcaaactt ggctcttat 
      cagtggttc accaatcag ggatatatc tgtgacagt 
      gataacagg actattttt cacaaaagt ctccttgct 
      agtgctttt ggaggggca gctggtgct tttgttgga 
      agtcctctc tatcaaatt accactcaa ctgcagtcg 
      aatgctgga gaggggatt gcggtggga catcagcat 
      aagcactct ggtcttgta caagctttc aaagcaata 
      tattcaagg catggaatg aaaggttta tggagaggt 
      tccaatgca aatgttata aggacagta gttggggct 
      tcagcacaa ctaacgact ttttccaaa actaaggac 
      atactaaga gagtacgaa ttctttcaa aggtctaca 
      acaatgact gcttttttc gcaagcatt gctggagga 
      atttgccag actgtcttt cagacgccc ttagattta 
      gtatgcatt agattaaat aatcagtat gtttacgcc 
      agtggacaa ggagcctta tatacagga atgactaat 
      tgtttgaca aaaattata caatctgaa gggtttttt 
      ggcttgtac aaaggtgct ggagttaat tatatgagg 
      atagctaga caaaccggt ttttgtcta atgttctgg 
      gaacttcta aaggattta catgtcaag cattttggt 
      ttaaataaa aacggacat tgtaatacg agtatatct 
      gatgaaata attaaattg taa
    mRNA sequence
    • >TC032499 RA
      tgaaacgaa taaaaattg caaggatca tctgaaaca 
      tacagagca ctaatgcct agctttaac catcgtcaa 
      cgatggact tcctaattg gcggtttag ctgcaatgg 
      gagccactt ttttcagca atccccttg acgttctta 
      aaactagaa tacagcttc aaggagaac tgaaacctc 
      gagggcaac atgaaatcc actacagac acgcactgc 
      atgctgctt atgttgtaa taaaacagg aaggaattt 
      ttgcattgc agaaaggac ttggagctg cacttctga 
      tgcatggag tgcgtaaca gtatcaaac ttggctctt 
      atcagtggt tcaccaatc agggatata tctgtgaca 
      gtgataaca ggactattt ttcacaaaa gtctccttg 
      ctagtgctt ttggagggg cagctggtg cttttgttg 
      gaagtcctc tctatcaaa ttaccactc aactgcagt 
      cgaatgctg gagagggga ttgcggtgg gacatcagc 
      ataagcact ctggtcttg tacaagctt tcaaagcaa 
      tatattcaa ggcatggaa tgaaaggtt tatggagag 
      gttccaatg caaatgtta taaggacag tagttgggg 
      cttcagcac aactaacga ctttttcca aaactaagg 
      acatactaa gagagtacg aattctttc aaaggtcta 
      caacaatga ctgcttttt tcgcaagca ttgctggag 
      gaatttgcc agactgtct ttcagacgc ccttagatt 
      tagtatgca ttagattaa ataatcagt atgtttacg 
      ccagtggac aaggagcct tatatacag gaatgacta 
      attgtttga caaaaatta tacaatctg aagggtttt 
      ttggcttgt acaaaggtg ctggagtta attatatga 
      ggatagcta gacaaaccg gtttttgtc taatgttct 
      gggaacttc taaaggatt tacatgtca agcattttg 
      gtttaaata aaaacggac attgtaata cgagtatat 
      ctgatgaaa taattaaat tgtaacttt ctgtttatt 
      tatttgagc ctagactag cattcgaat c
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY