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Gene information
General information
TC000442 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_seqid7: 5702693..5703467
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
    TC000442
    iBeetle screen iB_00094
    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)

    pupa eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 2 (3 dpi)
    Days post injection


    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: 11.11% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 11.11% (includes death as larva, prepupa, pupa, adult)

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


    Adult morphology
    motility irregular - (19 dpi)
    Days post injection

    Comment: animals avoid the middle of the vial and ca. 50% play dead, but are not

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC000442 RA
      MNRIFVFAA FVGAVFGQG YHGDPQTAA 
      ILNEQRYLS GDGKFGAQY SQEDGVQFK 
      EESDADGTR RGTYSYVDP NGQRHTVSY 
      TAGKHGFQA TGDGIPQPL PTTLPVAPQ 
      PQYTPLPQY NPPEYRDQY QPQYRPQPQ 
      YQPQPQYQP QPQYNPQQY QAQPQYRPQ 
      PVYTTTPAP HRFYPPGKL SLNRSPDGF 
      SYSFQKA
    CDS sequence
    • >TC000442 RA
      atgaacaga atattcgtc tttgctgct tttgtgggc 
      gctgttttc ggacagggc taccatggg gacccccaa 
      actgcggct atcctgaac gaacagagg tacctgagt 
      ggtgatggt aaattcggg gcccagtac tcccaagag 
      gatggggtg cagttcaag gaggagagt gacgctgat 
      ggcacccgt cgggggacc tactcctac gtggacccc 
      aacggccaa agacacacc gtgagctac actgctggc 
      aagcacggc ttccaagcc actggggat ggcatcccc 
      caaccctta ccgacgacc ttacccgtg gccccccag 
      ccccagtac acccccctt ccccagtac aaccccccg 
      gagtaccgc gaccagtac cagccccag tacaggccg 
      cagccccag taccagccc cagccccag taccaaccc 
      caaccccag tacaacccc cagcagtac caagcccag 
      ccacaatac aggccccag cccgtctac acgacgact 
      ccagcccct cacagattt taccctcct ggaaaactg 
      tcgcttaat aggtcgcca gatggcttc agttatagt 
      ttccagaag gcttaa
    mRNA sequence
    • >TC000442 RA
      ggcagtagt cgtagtttg cctctttgg tagaccatc 
      agttccact atcatgaac agaatattc gtctttgct 
      gcttttgtg ggcgctgtt ttcggacag ggctaccat 
      ggggacccc caaactgcg gctatcctg aacgaacag 
      aggtacctg agtggtgat ggtaaattc ggggcccag 
      tactcccaa gaggatggg gtgcagttc aaggaggag 
      agtgacgct gatggcacc cgtcggggg acctactcc 
      tacgtggac cccaacggc caaagacac accgtgagc 
      tacactgct ggcaagcac ggcttccaa gccactggg 
      gatggcatc ccccaaccc ttaccgacg accttaccc 
      gtggccccc cagccccag tacaccccc cttccccag 
      tacaacccc ccggagtac cgcgaccag taccagccc 
      cagtacagg ccgcagccc cagtaccag ccccagccc 
      cagtaccaa ccccaaccc cagtacaac ccccagcag 
      taccaagcc cagccacaa tacaggccc cagcccgtc 
      tacacgacg actccagcc cctcacaga ttttaccct 
      cctggaaaa ctgtcgctt aataggtcg ccagatggc 
      ttcagttat agtttccag aaggcttaa gtcgtagtt 
      agggatcat tgtcgcgta gcttaggtt cacaagtgt 
      ccattaatg tattatatt cataataaa atttttatt 
      taaat