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Gene information
General information
TC004326 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.

scaffold_seqid62: 254496..257764
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
    TC004326
    iBeetle screen iB_00679
    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: 20.0% (includes death as pupa, adult)

    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


    Fertility
    egg chamber not present - Number of animals showing the phenotype: 4

    tropharium partially structure fragmented - Number of animals showing the phenotype: 4

    Analysis of larval stage 1 cuticle
    remarks - cuticles: bad quality of preparation,
    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: 0.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)

    pupal molt delayed (still prepupa) - Number of animals showing the phenotype: 10 (11 dpi)
    Days post injection


    pupal molt not fulfilled (died as prepupa) - Number of animals showing the phenotype: 10 (16 dpi)
    Days post injection



    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC004326 RC
      MVIEKNPFP GNALEQFVL LSKSTKGAA CSELIKQVL 
      EAPGVYVFG ELLHMQNIE ELSNTPQKN FYNTLNLFA 
      YGTYKDYLA NKKDLIELS PAQKKKLQH LTIVTLATK 
      SKCIPYNTL LEELGITNV RDLEDLIIE AIYADIIHG 
      KLDQKNSQL EVDYAIGRD IRPEDINVI VNCLQDWCS 
      ACEGVLACV ETQIHRANN EKNKSMLRK AEVEQEISN 
      IKKTLKTQL QERSDGTDE AMATDNREA AAISEKGKK 
      STKIKGLKL LR
    CDS sequence
    • >TC004326 RC
      atggttatt gaaaaaaat cctttccct ggcaatgcc 
      ttggaacaa tttgtttta ttgtcaaag tcaacaaaa 
      ggggctgct tgttccgaa ctgatcaag caagtgttg 
      gaagccccc ggtgtttac gtttttggt gaattactt 
      cacatgcaa aacattgaa gagttgagt aatacacca 
      cagaaaaat ttctacaac actctcaac ttgtttgct 
      tacggaacc tacaaggat tatctagca aataagaag 
      gatttaatt gaattaagc ccagctcag aagaaaaag 
      cttcaacat ttaaccatt gttacgttg gccaccaaa 
      tcaaagtgt attccgtac aacaccctt ctggaagag 
      ctaggcata acaaatgtt cgtgatttg gaggactta 
      ataattgag gcgatttac gctgacatt attcatgga 
      aaattggac cagaaaaat agtcaattg gaggttgat 
      tatgccata ggtcgtgat attcgtcct gaagacatt 
      aatgtaatt gtgaattgt ctccaagac tggtgttcg 
      gcctgtgaa ggtgttttg gcttgtgtt gaaacacaa 
      atacataga gcaaataat gaaaaaaat aaatcaatg 
      ttgagaaaa gctgaagtg gagcaagag attagtaac 
      attaaaaag actctgaaa actcaatta caagaacgc 
      tctgatgga actgacgag gctatggct actgataac 
      agggaagct gcagcaatt agcgaaaaa ggaaaaaag 
      tcaactaaa atcaaagga ttaaaactt cttcgt
    mRNA sequence
    • >TC004326 RC
      cgtgtgaca gctgatcgg ttgttgtgt ttaaaaagt 
      gcttcaaaa acaacaaat taactaata aaaactaaa 
      aaaatggtt attgaaaaa aatcctttc cctggcaat 
      gccttggaa caatttgtt ttattgtca aagtcaaca 
      aaaggggct gcttgttcc gaactgatc aagcaagtg 
      ttggaagcc cccggtgtt tacgttttt ggtgaatta 
      cttcacatg caaaacatt gaagagttg agtaataca 
      ccacagaaa aatttctac aacactctc aacttgttt 
      gcttacgga acctacaag gattatcta gcaaataag 
      aaggattta attgaatta agcccagct cagaagaaa 
      aagcttcaa catttaacc attgttacg ttggccacc 
      aaatcaaag tgtattccg tacaacacc cttctggaa 
      gagctaggc ataacaaat gttcgtgat ttggaggac 
      ttaataatt gaggcgatt tacgctgac attattcat 
      ggaaaattg gaccagaaa aatagtcaa ttggaggtt 
      gattatgcc ataggtcgt gatattcgt cctgaagac 
      attaatgta attgtgaat tgtctccaa gactggtgt 
      tcggcctgt gaaggtgtt ttggcttgt gttgaaaca 
      caaatacat agagcaaat aatgaaaaa aataaatca 
      atgttgaga aaagctgaa gtggagcaa gagattagt 
      aacattaaa aagactctg aaaactcaa ttacaagaa 
      cgctctgat ggaactgac gaggctatg gctactgat 
      aacagggaa gctgcagca attagcgaa aaaggaaaa 
      aagtcaact aaaatcaaa ggattaaaa cttcttcgt 
      tagtgatgt cttccaagt tgggatcac accgccagt 
      tggtacaca aataatgtt taattgctg attttcttg 
      taaatttgt atttgtggt taataaaaa attttttct 
      tac