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Gene information
General information
TC014489 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_007420.3: 11150233..11151198
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
    TC014489
    iBeetle screen iB_10087

    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: 10.0% (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


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 embryo/egg not developed/not fertilized - Penetrance (percentage of animals/eggs on slide): more than 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): more than 80%
    Comment: some wildtype larvae present. n=4
    cuticle remnants strongly fragmented (cuticle crumbs) - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    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
    • >TC014489 RA
      MPLPSALAA RLAKRGLLA GKKSDDQTK KRNFKGWST 
      CPNKSNIFH ECTYWCETH WKGVLTPDP RYMRNMQKL 
      LFKYPLPNS WTEVFDKGL GRYYYWNME NDLVSWLPP 
      RHPKSVKTL SAAKLREKR VLAAENDKS DKGSDDNDS 
      DDDDRGHKH SDRRDRDRD RDDRRHRKE DRHGRKKRR 
      EDIIDPMDP AAYSDIPQG TWSDGLESN KTRADNTAS 
      GVLYQQRPY PAPGEVLAN NNKEKSKKK
    CDS sequence
    • >TC014489 RA
      atgcctctc ccttcggct ctagcggcc cgtttggca 
      aaaaggggg ctgttggct ggcaaaaaa tcggacgat 
      cagacgaaa aaacgcaac ttcaaaggt tggtccacc 
      tgccccaac aaaagcaac attttccac gaatgcacc 
      tactggtgt gagacgcac tggaagggt gttctgacc 
      cccgacccc cgttacatg cgaaacatg caaaaactc 
      ctcttcaaa taccccttg cccaacagt tggactgag 
      gtctttgac aagggattg gggcgttac tactactgg 
      aacatggaa aatgacttg gtttcgtgg ctgcctcca 
      agacaccct aaatcagtg aagacctta agtgcggcg 
      aaacttcgc gagaaacgg gttctagcc gcggaaaat 
      gacaagtct gataaggga tcggacgat aatgactcg 
      gatgatgat gacaggggt cataaacat tcggatcgg 
      agggacagg gaccgggat agggatgac aggaggcac 
      aggaaggag gacagacac gggaggaag aaacgaagg 
      gaggatatc atagacccc atggaccca gcggcttat 
      tcggacatc ccccaaggg acgtggagt gacggcctg 
      gagagcaat aaaacaagg gctgataat acggcttct 
      ggagtcctc taccagcaa agaccttac cccgctcct 
      ggagaagtc ttagctaat aataataaa gaaaaaagc 
      aagaaaaaa taa
    mRNA sequence
    • >TC014489 RA
      ggataaaaa atccgaatc cggccctgt gctgtcacc 
      ctcgacagt tgaggttat gtaacattg aggctatat 
      ttaccaaac aaatcgcta aaatgcctc tcccttcgg 
      ctctagcgg cccgtttgg caaaaaggg ggctgttgg 
      ctggcaaaa aatcggacg atcagacga aaaaacgca 
      acttcaaag gttggtcca cctgcccca acaaaagca 
      acattttcc acgaatgca cctactggt gtgagacgc 
      actggaagg gtgttctga cccccgacc cccgttaca 
      tgcgaaaca tgcaaaaac tcctcttca aatacccct 
      tgcccaaca gttggactg aggtctttg acaagggat 
      tggggcgtt actactact ggaacatgg aaaatgact 
      tggtttcgt ggctgcctc caagacacc ctaaatcag 
      tgaagacct taagtgcgg cgaaacttc gcgagaaac 
      gggttctag ccgcggaaa atgacaagt ctgataagg 
      gatcggacg ataatgact cggatgatg atgacaggg 
      gtcataaac attcggatc ggagggaca gggaccggg 
      atagggatg acaggaggc acaggaagg aggacagac 
      acgggagga agaaacgaa gggaggata tcatagacc 
      ccatggacc cagcggctt attcggaca tcccccaag 
      ggacgtgga gtgacggcc tggagagca ataaaacaa 
      gggctgata atacggctt ctggagtcc tctaccagc 
      aaagacctt accccgctc ctggagaag tcttagcta 
      ataataata aagaaaaaa gcaagaaaa aataattgt 
      gtattagtt tattatcaa tatatcgct aaaaccatt
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY