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Gene information
General information
TC000028 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: 15862753..15867130
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
    TC000028
    iBeetle screen iB_02965

    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: 20.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 2 (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): 30% - 50%

    larval musculature fluorescence decreased - Penetrance (percentage of animals/eggs on slide): less than 30%

    Comment: Potentially due to death.
    larval musculature irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    remarks: muscle pattern potentially obscured by segmentation defect,
    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): 50% - 80%
    larva circumference irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    abdomen segment number decreased - 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
    • >TC000028 RA
      MPPKAKPAA PSKKTEQKK KEKVIEDKT FGLKNKKGA 
      KQQKFIQQV EKQVKTGGH HPVKEDGGK KAEKEKKLK 
      EQKEMAILF KTVQTQKVE KGTDPKSVV CSFFKQGQC 
      GKGDKCKFS HDLSIERKA EKRSLYVDM RDDEEETME 
      NWDEQKLFE VIEKKHGKT GKMPTTDII CKHFLEAVE 
      KSKYGWFWQ CPSGESCIY RHALPPGFT LKKDKKKDD 
      KRDEISLED LIEKERAAL GSTQTKVTL ETFLAWKKR 
      KIQEKKDAA KRDEDKKRT DFKAGKQVG LSGREMFSF 
      NPELAAQNN MDEGDEAFD SYNYSDTED DEADYKEIN 
      LDLISAEEV DGTGTVASE DRLKTSSKV TENGEEGAS 
      AVPINENLF LEEDLEALD EELNDLDLE E
    CDS sequence
    • >TC000028 RA
      atgccaccg aaggccaaa ccggcagca cccagtaag 
      aaaactgaa cagaaaaag aaagaaaaa gtgattgag 
      gacaagact ttcggtttg aaaaataaa aaaggggcc 
      aagcagcag aaattcata cagcaagtc gagaaacaa 
      gttaaaact gggggtcac catcctgtt aaagaagat 
      ggggggaag aaagcggaa aaggagaag aagttgaag 
      gaacagaag gaaatggct attttattt aagaccgtg 
      caaacgcag aaagtcgag aaagggact gatcccaag 
      tctgtggtt tgttcgttc ttcaaacag ggccagtgc 
      ggcaaaggt gataagtgc aaattttcc catgatttg 
      agtatagag cggaaggcg gagaaaagg tcgctgtat 
      gtggatatg agagatgat gaagaagaa actatggaa 
      aactgggat gagcaaaag ctgtttgaa gtgatagag 
      aagaaacat ggaaagacg gggaaaatg cccactact 
      gacattatt tgtaaacat ttcttggaa gctgtagaa 
      aaatcaaag tatggctgg ttctggcaa tgtccctct 
      ggagaaagc tgtatttac agacacgct ctcccacca 
      ggtttcacg ttaaagaaa gataagaaa aaagacgac 
      aaacgtgat gaaatatca ctggaagat ctaattgaa 
      aaggaaaga gcagcacta ggttctact caaactaaa 
      gtcacgtta gagacattt ttagcgtgg aaaaaacgt 
      aaaattcag gaaaaaaaa gacgccgca aaacgggat 
      gaagacaaa aaacgaacc gactttaag gcaggcaaa 
      caagtcgga cttagtggt agagaaatg tttagtttt 
      aatcctgaa ttggcggcg cagaataac atggatgaa 
      ggcgatgaa gcgtttgat tcttacaat tattcagac 
      accgaagat gatgaagcg gattataaa gaaatcaat 
      ttagatcta attagtgcc gaagaggtt gacgggact 
      ggaactgtg gcctctgaa gaccggctg aagacaagc 
      tcaaaagta acagaaaat ggtgaagaa ggtgcatct 
      gcagtaccc atcaatgaa aatttgttc cttgaggag 
      gatttagaa gctttagat gaagaacta aacgactta 
      gatttagaa gaataa
    mRNA sequence
    • >TC000028 RA
      tctgtcaaa tccccgacc tgtcactgt gcatgtgga 
      acttccgaa agcgaaata tttttcgtt ttctttgaa 
      aaaattatc aagattatt taattctct caattagaa 
      ctgtaaaaa atgccaccg aaggccaaa ccggcagca 
      cccagtaag aaaactgaa cagaaaaag aaagaaaaa 
      gtgattgag gacaagact ttcggtttg aaaaataaa 
      aaaggggcc aagcagcag aaattcata cagcaagtc 
      gagaaacaa gttaaaact gggggtcac catcctgtt 
      aaagaagat ggggggaag aaagcggaa aaggagaag 
      aagttgaag gaacagaag gaaatggct attttattt 
      aagaccgtg caaacgcag aaagtcgag aaagggact 
      gatcccaag tctgtggtt tgttcgttc ttcaaacag 
      ggccagtgc ggcaaaggt gataagtgc aaattttcc 
      catgatttg agtatagag cggaaggcg gagaaaagg 
      tcgctgtat gtggatatg agagatgat gaagaagaa 
      actatggaa aactgggat gagcaaaag ctgtttgaa 
      gtgatagag aagaaacat ggaaagacg gggaaaatg 
      cccactact gacattatt tgtaaacat ttcttggaa 
      gctgtagaa aaatcaaag tatggctgg ttctggcaa 
      tgtccctct ggagaaagc tgtatttac agacacgct 
      ctcccacca ggtttcacg ttaaagaaa gataagaaa 
      aaagacgac aaacgtgat gaaatatca ctggaagat 
      ctaattgaa aaggaaaga gcagcacta ggttctact 
      caaactaaa gtcacgtta gagacattt ttagcgtgg 
      aaaaaacgt aaaattcag gaaaaaaaa gacgccgca 
      aaacgggat gaagacaaa aaacgaacc gactttaag 
      gcaggcaaa caagtcgga cttagtggt agagaaatg 
      tttagtttt aatcctgaa ttggcggcg cagaataac 
      atggatgaa ggcgatgaa gcgtttgat tcttacaat 
      tattcagac accgaagat gatgaagcg gattataaa 
      gaaatcaat ttagatcta attagtgcc gaagaggtt 
      gacgggact ggaactgtg gcctctgaa gaccggctg 
      aagacaagc tcaaaagta acagaaaat ggtgaagaa 
      ggtgcatct gcagtaccc atcaatgaa aatttgttc 
      cttgaggag gatttagaa gctttagat gaagaacta 
      aacgactta gatttagaa gaataaggg tgctaatat 
      ttagcattg ttggaattt aatttcatt actaaactc 
      gtaatgcac accagagat caacggtt
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY