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Gene information
General information
TC012299 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: 3887772..3890028
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
    TC012299
    iBeetle screen iB_01964

    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: 40.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 4 (3 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 embryo/egg with embryonic tissue potentially present, no cuticle, no fluorescence - 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%
    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
    • >TC012299 RA
      MDLVTQTSL EALLLKATN SQNQNIDTA AVEAFCALV 
      NKEKDGAHI GVKVIANRL PSGNEKELL QTLNILDTC 
      MSKCGTAFQ SEVGKFRFL NEMIKLVSP KYLGSQTPL 
      VVKQKVLQL MYIWTLDYP KETKIKEAY DMLRKQGVI 
      KEIPNPNIV QNANIPRKK AANSVFQDE EKSKILQKL 
      LQSKDPEDI QAANWLIKS MVKEDDKRA ELKSRRVSE 
      LESVQNNVR LLNEMLDSY KPGISSADE LDLIKELHQ 
      SCERLRPNL MKLTAETQQ NEEILGDVL QASDELSQV 
      FGKYNRIIV QGVSVVDDG SNALLDFEG ADEAQCRNS 
      DTKEIDVLC DIFNASNIP DSADILKPV SVLTCDKNE 
      NGKEVNKFK ALEELDVLS EHFIKENLP TSRQGTFNN 
      RQDKIPMNL LTKLSETKP VQSNGKNES ISESLSLDL 
      SYLISKPKD EPKTNTMEN STDDCLVDI ADEKMLEAG 
      ETPQRKPSL TDPAKNEIK LNDIFIKLE DIKPSPVPP 
      LTILNERNG ITITLHFVK DQPRDDVHV FVVTTINKN 
      EMPLTNYLF QAVVPKGCK VKLQPPSAT TLPAYNPFL 
      PPSAITQIM LIANPDDVQ VFLKFFVSF VMDEETCTE 
      IGEVETLPM
    CDS sequence
    • >TC012299 RA
      atggatttg gtgactcaa actagtctc gaagccctg 
      cttttaaaa gccacaaac tcccaaaac caaaacatt 
      gacaccgcc gctgttgag gccttctgc gcccttgta 
      aacaaagag aaagatggg gcacacatc ggggttaaa 
      gtgatagca aaccggctt ccttcaggt aacgagaaa 
      gagctgctc caaacgctc aacatcctg gatacgtgc 
      atgagcaaa tgtggcaca gctttccag tcagaagtc 
      ggcaaattc cggtttctg aacgaaatg attaagcta 
      gtttcgccc aaatatctg gggtcgcag acgccgctc 
      gtggtgaag caaaaagtg ctgcaactg atgtacatt 
      tggacactg gactacccc aaagaaacg aagataaag 
      gaagcgtac gatatgctg cgaaagcag ggcgttatc 
      aaggaaatc ccgaacccc aacatagta caaaacgcc 
      aacattccg aggaaaaag gccgcgaat tccgtgttc 
      caagacgag gagaaatca aaaattttg caaaagctg 
      ctgcaaagt aaagacccc gaggacatc caagccgcc 
      aattggctc attaaaagt atggtcaag gaggacgat 
      aaaagggcg gaattgaag agccggcgc gtttccgag 
      cttgagtca gtacaaaac aatgtgaga ttattgaat 
      gaaatgctc gattcgtac aaaccgggc atttcttcc 
      gccgatgag ctggacctt attaaagag ctgcatcaa 
      agttgtgag cgtctgaga ccaaattta atgaaacta 
      accgcagag acacaacaa aatgaagaa atcttaggt 
      gatgttttg caagcaagc gatgagttg agccaagtt 
      tttggcaag tataacagg attattgtg cagggagtt 
      agtgtcgtg gacgatggc agcaatgcc ttgcttgat 
      tttgagggc gcagacgag gctcagtgt cggaactct 
      gataccaaa gaaattgac gttttatgt gatattttc 
      aacgcttca aacattccg gactctgct gatattttg 
      aaacctgtt tcagtatta acttgtgat aaaaacgaa 
      aatgggaaa gaggtgaat aaattcaaa gccttggaa 
      gagttggac gttttgagc gaacacttt attaaggaa 
      aatctccca actagcaga caaggcact tttaataat 
      aggcaagac aaaatccca atgaatctc cttacaaag 
      ctatctgag acgaaacca gttcagagt aacggaaag 
      aacgaatcg attagtgaa tctctaagt ctagatttg 
      agttattta ataagcaaa ccaaaagac gaaccaaaa 
      acaaacaca atggaaaat tccactgat gactgtttg 
      gtggacata gctgacgag aaaatgctg gaagctggg 
      gaaaccccc caacgaaag cccagctta actgacccg 
      gcaaaaaat gaaataaaa ctgaacgac atttttatc 
      aaattggaa gatatcaag ccaagtccc gtcccacct 
      ctcacgatt ttaaacgag aggaacgga atcacaata 
      accctgcat ttcgtgaaa gaccagccc cgagacgac 
      gtgcatgtc tttgttgtg accacaatt aacaaaaac 
      gagatgcct ttgactaac tacctgttc caggctgtg 
      gtacctaag ggctgcaaa gtcaaactg caacctccc 
      tcagcaacc accctcccg gcttacaac ccgttcctg 
      cccccttct gccataacg caaatcatg ctgatcgca 
      aatcctgat gatgtacaa gtttttctc aagtttttt 
      gtaagtttt gtaatggac gaagagact tgtactgaa 
      atcggcgag gttgaaact cttccaatg taa
    mRNA sequence
    • >TC012299 RA
      tgacgttta agtgacatg aaaataaaa tgattgaaa 
      caatttaaa aacccataa aatggattt ggtgactca 
      aactagtct cgaagccct gcttttaaa agccacaaa 
      ctcccaaaa ccaaaacat tgacaccgc cgctgttga 
      ggccttctg cgcccttgt aaacaaaga gaaagatgg 
      ggcacacat cggggttaa agtgatagc aaaccggct 
      tccttcagg taacgagaa agagctgct ccaaacgct 
      caacatcct ggatacgtg catgagcaa atgtggcac 
      agctttcca gtcagaagt cggcaaatt ccggtttct 
      gaacgaaat gattaagct agtttcgcc caaatatct 
      ggggtcgca gacgccgct cgtggtgaa gcaaaaagt 
      gctgcaact gatgtacat ttggacact ggactaccc 
      caaagaaac gaagataaa ggaagcgta cgatatgct 
      gcgaaagca gggcgttat caaggaaat cccgaaccc 
      caacatagt acaaaacgc caacattcc gaggaaaaa 
      ggccgcgaa ttccgtgtt ccaagacga ggagaaatc 
      aaaaatttt gcaaaagct gctgcaaag taaagaccc 
      cgaggacat ccaagccgc caattggct cattaaaag 
      tatggtcaa ggaggacga taaaagggc ggaattgaa 
      gagccggcg cgtttccga gcttgagtc agtacaaaa 
      caatgtgag attattgaa tgaaatgct cgattcgta 
      caaaccggg catttcttc cgccgatga gctggacct 
      tattaaaga gctgcatca aagttgtga gcgtctgag 
      accaaattt aatgaaact aaccgcaga gacacaaca 
      aaatgaaga aatcttagg tgatgtttt gcaagcaag 
      cgatgagtt gagccaagt ttttggcaa gtataacag 
      gattattgt gcagggagt tagtgtcgt ggacgatgg 
      cagcaatgc cttgcttga ttttgaggg cgcagacga 
      ggctcagtg tcggaactc tgataccaa agaaattga 
      cgttttatg tgatatttt caacgcttc aaacattcc 
      ggactctgc tgatatttt gaaacctgt ttcagtatt 
      aacttgtga taaaaacga aaatgggaa agaggtgaa 
      taaattcaa agccttgga agagttgga cgttttgag 
      cgaacactt tattaagga aaatctccc aactagcag 
      acaaggcac ttttaataa taggcaaga caaaatccc 
      aatgaatct ccttacaaa gctatctga gacgaaacc 
      agttcagag taacggaaa gaacgaatc gattagtga 
      atctctaag tctagattt gagttattt aataagcaa 
      accaaaaga cgaaccaaa aacaaacac aatggaaaa 
      ttccactga tgactgttt ggtggacat agctgacga 
      gaaaatgct ggaagctgg ggaaacccc ccaacgaaa 
      gcccagctt aactgaccc ggcaaaaaa tgaaataaa 
      actgaacga catttttat caaattgga agatatcaa 
      gccaagtcc cgtcccacc tctcacgat tttaaacga 
      gaggaacgg aatcacaat aaccctgca tttcgtgaa 
      agaccagcc ccgagacga cgtgcatgt ctttgttgt 
      gaccacaat taacaaaaa cgagatgcc tttgactaa 
      ctacctgtt ccaggctgt ggtacctaa gggctgcaa 
      agtcaaact gcaacctcc ctcagcaac caccctccc 
      ggcttacaa cccgttcct gcccccttc tgccataac 
      gcaaatcat gctgatcgc aaatcctga tgatgtaca 
      agtttttct caagttttt tgtaagttt tgtaatgga 
      cgaagagac ttgtactga aatcggcga ggttgaaac 
      tcttccaat gtaaagcat tttaatttt agtgtaatt 
      aaatcgtgc ttaagtca
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY