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Gene information
General information
TC012528 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_007424.3: 11022925..11026789
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
    TC012528
    iBeetle screen iB_05259

    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)

    pupa eclosion not fulfilled (died as pupa) - 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
    mature egg deposition blocked number 1st lateral oviduct: 3 number 2nd lateral oviduct: 2 - Number of animals showing the phenotype: 1
    blocked number 1st lateral oviduct: 3 - Number of animals showing the phenotype: 1
    yolk structure irregular - Number of animals showing the phenotype: 4

    Comment: The yolk in the early vitellogenic egg chambers looks slightly irregular.
    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%

    Comment: yolk cells look slightly unusual. They appear to be a bit too transparent.
    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
    egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): more than 80%
    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: 10.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 10.0% (includes death as larva, prepupa, pupa, adult)

    Fertility
    egg production: number no/almost no eggs -
    mature egg slightly size decreased - Number of animals showing the phenotype: 4

    yolk sometimes mostlynot present - Number of animals showing the phenotype: 4
    yolk structure irregular - Number of animals showing the phenotype: 4
    Comment: Yolk seems to be more granular than usual
    yolk shiny -
    Comment: not as opaque in lateral illumination as usual
    fertility after larval injection decreased (no living offspring) -
    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC012528 RA
      MVANNTTSA FLQTLIFFM SQSNFQQDT SYNYYFLEI 
      GKTNNASKC YGIYGCFQL SPPWTSENR PVSLFPEDL 
      TKIEPRYPL YTRKNPTKP TYIDLNEYD VIGSTGINP 
      RNPVYVISH GFMEGGGIY WIVDMAQKL LKIHDCSVI 
      VVDWQGGSG PPYTQAVAN IRLVGAMTA HLLHDLWKH 
      IPDMNLDHV HCIGHSLGA HLCGYVGYT LHRDFKLTL 
      GRITGLDPA EPHFAKAQP PVRLDRTAA KYVDVVHTD 
      ASQFIRGGL GMTERIGHV DYYPNGGTN QPGCGKSIA 
      KYIDEANGS FFLGVRKYM GCNHMRSYE YFIESINPN 
      RACSFLTVG CNNYADFLA GKCFDCGRR NQKCIQFGY 
      YSHKEYKEL VKRRVISPD SNMVQFLIT GEAKPYCRG 
      HYRITVKIS NSGESLQHG GEVGQLFFT MHETTDGKG 
      PKTHTVPLN KGGFHEPGK IYTAVVPTS EVKKIKAVE 
      VEWRYQSSY LNPLTWRLL TVPKIYIEK ITIDALEIR 
      QSLTVCPKD GQALHTSQP HFMIPSYC
    CDS sequence
    • >TC012528 RA
      atggtcgca aataacacc acttctgct ttcttacaa 
      acccttatc tttttcatg agccaaagc aacttccag 
      caagacacc tcctacaac tattacttt ttagaaatt 
      gggaaaacc aacaatgcg tccaaatgt tacggaatc 
      tacggctgc ttccagctg tccccccca tggacctcg 
      gagaaccgc cccgtgtcc ctgttcccg gaagacctg 
      actaagatc gagccgcgg tacccgctc tacacgcgc 
      aaaaacccc accaaaccc acctacatc gacttgaat 
      gagtacgac gtgatcggc tcgacgggc atcaacccc 
      cgcaacccc gtctacgtg atttcgcac ggcttcatg 
      gagggtgga gggatatat tggattgta gacatggcc 
      cagaagtta ttgaaaatt cacgactgt tccgtcatc 
      gtggtcgac tggcaaggg ggctcaggg cccccctac 
      acacaagca gtcgccaat attcggcta gtgggggca 
      atgactgcg catcttttg cacgactta tggaagcac 
      atccctgat atgaacctg gaccacgtg cactgcatc 
      ggacactcg ctgggggcc cacttgtgc ggatacgtg 
      ggatatacc ttgcaccgg gactttaaa ttgactttg 
      gggcgcata accggcctc gacccggct gaaccgcac 
      tttgcaaag gcccaaccg ccggttcgg ttagaccga 
      accgccgcc aaatacgtc gatgtggtt cacaccgac 
      gctagtcag tttattcga ggaggacta ggtatgact 
      gaaaggatc ggtcacgtg gactattac ccaaatggg 
      ggcacaaac cagcccggc tgcggcaaa agcatagcc 
      aaatacatc gatgaggcc aatgggagc tttttcttg 
      ggtgtgagg aaatacatg ggctgcaat cacatgcga 
      agttacgaa tatttcatt gaaagcatt aatcccaac 
      agggcgtgt tcgtttcta acagttgga tgcaacaac 
      tacgcggat tttttggcg ggaaaatgt ttcgattgc 
      ggccgaaga aaccaaaaa tgcatccag ttcggctac 
      tacagccac aaagaatac aaagaactg gtaaaacgc 
      cgggtaatt tcacccgac tcgaacatg gtccagttt 
      ttgataaca ggagaagcg aaaccatac tgccgcggg 
      cattacaga ataacagta aaaatttca aactctggc 
      gaaagcctc caacatggt ggcgaagtg ggccaactg 
      ttcttcaca atgcatgaa acaaccgac ggtaaaggg 
      ccgaaaacg cacacggtg cccttaaac aagggcggg 
      ttccacgag cccgggaaa atctacaca gcggtggtg 
      ccaacgtcc gaagtcaag aaaatcaaa gctgttgaa 
      gtcgaatgg cggtaccag agcagctac ttgaaccct 
      ttgacctgg aggctcttg accgtcccg aaaatatac 
      atcgagaaa ataaccatc gacgctctg gaaatcagg 
      cagagtttg actgtgtgt ccgaaagac ggccaggcg 
      ttacacacc tcgcaaccg cacttcatg atcccgtct 
      tattgttaa
    mRNA sequence
    • >TC012528 RA
      ttccgttat aatcagttc tttcacaaa gtgcgccca 
      agtcattac caaaaaatc acaatggtc gcaaataac 
      accacttct gctttctta caaaccctt atctttttc 
      atgagccaa agcaacttc cagcaagac acctcctac 
      aactattac tttttagaa attgggaaa accaacaat 
      gcgtccaaa tgttacgga atctacggc tgcttccag 
      ctgtccccc ccatggacc tcggagaac cgccccgtg 
      tccctgttc ccggaagac ctgactaag atcgagccg 
      cggtacccg ctctacacg cgcaaaaac cccaccaaa 
      cccacctac atcgacttg aatgagtac gacgtgatc 
      ggctcgacg ggcatcaac ccccgcaac cccgtctac 
      gtgatttcg cacggcttc atggagggt ggagggata 
      tattggatt gtagacatg gcccagaag ttattgaaa 
      attcacgac tgttccgtc atcgtggtc gactggcaa 
      gggggctca gggcccccc tacacacaa gcagtcgcc 
      aatattcgg ctagtgggg gcaatgact gcgcatctt 
      ttgcacgac ttatggaag cacatccct gatatgaac 
      ctggaccac gtgcactgc atcggacac tcgctgggg 
      gcccacttg tgcggatac gtgggatat accttgcac 
      cgggacttt aaattgact ttggggcgc ataaccggc 
      ctcgacccg gctgaaccg cactttgca aaggcccaa 
      ccgccggtt cggttagac cgaaccgcc gccaaatac 
      gtcgatgtg gttcacacc gacgctagt cagtttatt 
      cgaggagga ctaggtatg actgaaagg atcggtcac 
      gtggactat tacccaaat gggggcaca aaccagccc 
      ggctgcggc aaaagcata gccaaatac atcgatgag 
      gccaatggg agctttttc ttgggtgtg aggaaatac 
      atgggctgc aatcacatg cgaagttac gaatatttc 
      attgaaagc attaatccc aacagggcg tgttcgttt 
      ctaacagtt ggatgcaac aactacgcg gattttttg 
      gcgggaaaa tgtttcgat tgcggccga agaaaccaa 
      aaatgcatc cagttcggc tactacagc cacaaagaa 
      tacaaagaa ctggtaaaa cgccgggta atttcaccc 
      gactcgaac atggtccag tttttgata acaggagaa 
      gcgaaacca tactgccgc gggcattac agaataaca 
      gtaaaaatt tcaaactct ggcgaaagc ctccaacat 
      ggtggcgaa gtgggccaa ctgttcttc acaatgcat 
      gaaacaacc gacggtaaa gggccgaaa acgcacacg 
      gtgccctta aacaagggc gggttccac gagcccggg 
      aaaatctac acagcggtg gtgccaacg tccgaagtc 
      aagaaaatc aaagctgtt gaagtcgaa tggcggtac 
      cagagcagc tacttgaac cctttgacc tggaggctc 
      ttgaccgtc ccgaaaata tacatcgag aaaataacc 
      atcgacgct ctggaaatc aggcagagt ttgactgtg 
      tgtccgaaa gacggccag gcgttacac acctcgcaa 
      ccgcacttc atgatcccg tcttattgt taatcgtac 
      ctgcgtctg tatttatta cataaacca ttaaaaaca
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY