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Gene information
General information
TC030304 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: 3525572..3526945
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
    TC030304
    iBeetle screen iB_07596

    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 with embryonic tissue, no cuticle, but muscle fluorescence - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): less than 30%
    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): 30% - 50%
    head shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    head not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    thorax shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    thorax not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    abdomen shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    abdomen not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    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
    • >TC030304 RA
      MRRLKKLLL VDIRQKLFK PRNRQQVVM VQSRVNFWK 
      KIYFMFTGM GVATMFFWA LFPIMDGTV KEHRLPFLA 
      WYPFSVNKS PFYEITYIY QIVSVFFIV IVNMNSDML 
      LVALMNILG VQCDLLCDN LKNIQFRER INEEFLRCV 
      NHHMQILSY ASDCNKFFN TIVLAQFFT TVVSLGLTM 
      YQLTIVTPF TSEFYSFIV YGGAVLMEI FLYCWFGNE 
      VEFKSLNIP FASFGFDWT IGSVGLQKN LIIFIAKSQ 
      RPIRMSALN LFHLSLETF VKILRTAYS YFALLNNVN 
      SLN
    CDS sequence
    • >TC030304 RA
      atgagaagg ctcaagaaa cttctgttg gtcgacata 
      agacaaaaa ctgtttaaa ccaagaaac cgtcaacaa 
      gtagttatg gtgcaaagc agggtgaat ttttggaag 
      aagatttat tttatgttt actggaatg ggtgttgcg 
      actatgttt ttctgggct cttttccct attatggat 
      gggacggtt aaagaacac cgattgcct tttttggca 
      tggtatccc ttcagtgtc aataaatca ccgttttat 
      gaaatcact tatatttat caaattgtt agcgttttc 
      ttcattgta atcgtaaac atgaactcg gacatgttg 
      ttggttgct ttaatgaat attctgggg gtgcagtgt 
      gacttgttg tgtgataat ttgaaaaat atacaattt 
      agggaaaga ataaatgaa gagtttctt cggtgcgtt 
      aatcatcat atgcaaatt ttaagctac gcctccgat 
      tgcaacaag tttttcaac acaatcgtc ttagcccaa 
      tttttcacg actgtggtc tctttggga ctgacaatg 
      tatcagttg acaattgtt acccctttt accagtgaa 
      ttttactcg tttatagtc tatggaggt gcggtttta 
      atggaaata tttttatac tgctggttt ggaaatgaa 
      gttgaattt aaaagcttg aacattcct ttcgcctcg 
      tttgggttt gactggact attggtagc gtcgggttg 
      caaaaaaat ttaattatt ttcattgca aaaagtcaa 
      cgaccaatt agaatgagt gcccttaac ttgtttcat 
      ctgtcacta gaaactttc gttaaaatt ttacgtacg 
      gcttactcg tattttgcc cttttaaac aatgtcaat 
      tcattaaat tag
    mRNA sequence
    • >TC030304 RA
      tggaagaag acttcgact ttttatcct cccttcaaa 
      ctacccttt tctgcctaa ggtgtgttg gaacgtggc 
      cttccaaca catataaac tggatgcct acactttat 
      atgccactg catccataa caatttgtt tgttcggcc 
      acaattttt ttcaaaccg tcaacatct tcttcattt 
      tcaacgatc taaacaccc tcactggtg tcatcttcg 
      tcgcactta cttgcttgg tagcaattt taaaatcac 
      tgctattta ttttcaata tgagaaggc tcaagaaac 
      ttctgttgg tcgacataa gacaaaaac tgtttaaac 
      caagaaacc gtcaacaag tagttatgg tgcaaagca 
      gggtgaatt tttggaaga agatttatt ttatgttta 
      ctggaatgg gtgttgcga ctatgtttt tctgggctc 
      ttttcccta ttatggatg ggacggtta aagaacacc 
      gattgcctt ttttggcat ggtatccct tcagtgtca 
      ataaatcac cgttttatg aaatcactt atatttatc 
      aaattgtta gcgttttct tcattgtaa tcgtaaaca 
      tgaactcgg acatgttgt tggttgctt taatgaata 
      ttctggggg tgcagtgtg acttgttgt gtgataatt 
      tgaaaaata tacaattta gggaaagaa taaatgaag 
      agtttcttc ggtgcgtta atcatcata tgcaaattt 
      taagctacg cctccgatt gcaacaagt ttttcaaca 
      caatcgtct tagcccaat ttttcacga ctgtggtct 
      ctttgggac tgacaatgt atcagttga caattgtta 
      cccctttta ccagtgaat tttactcgt ttatagtct 
      atggaggtg cggttttaa tggaaatat ttttatact 
      gctggtttg gaaatgaag ttgaattta aaagcttga 
      acattcctt tcgcctcgt ttgggtttg actggacta 
      ttggtagcg tcgggttgc aaaaaaatt taattattt 
      tcattgcaa aaagtcaac gaccaatta gaatgagtg 
      cccttaact tgtttcatc tgtcactag aaactttcg 
      ttaaaattt tacgtacgg cttactcgt attttgccc 
      ttttaaaca atgtcaatt cattaaatt agctttttt 
      cataatatt taagtatta ataattaat gtagattta 
      t
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