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Gene information
General information
TC030232 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_007425.3: 2137622..2139051
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
    TC030232
    iBeetle screen iB_08338

    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)

    adult lethality - Number of animals showing the phenotype: 2 (9 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): less than 30%

    embryo/egg with muscle fluorescence/pattern, but not hatched (pot. late embryonic lethality) - Penetrance (percentage of animals/eggs on slide): 30% - 50%

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

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    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): 30% - 50%
    head not present - Penetrance (percentage of animals/eggs on slide): less than 30%

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

    thorax not present - Penetrance (percentage of animals/eggs on slide): less than 30%
    thorax shape irregular - Penetrance (percentage of animals/eggs on slide): less than 30%
    abdomen not present - 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
    • >TC030232 RA
      MQALTLTSI FAPQHVVSK ILGVLPFTI QDGRPVFSK 
      LGLCHSFLC IILYVFVAQ SYYQTIFDD TTSGFITLK 
      ILKFITIAR YVGYTAVMI NTFVAVGLT YKKFVALLS 
      KLYGIDRGL GKLGLETFV VASGYRIKK KMFLLVILE 
      YLIGNIVSI FSSQFNDIS GAFVLLYPR MIGCNLSVT 
      YFGTTLLLE ERFKLVNKF LLGECFKSR NCTKYPTDP 
      NFCDKIKLS VNIHKELTR ICRAINSIF SVFFLLWVT 
      VNFLCLVSD LYIVAYLIS SGQFFNYWQ GELKAFKNV 
      CIYSIEMFY IARRSSRLC SQANRTKTI LVGLKVDLS 
      KEEERNTII TSSLELRQF TLQITACRL FTIDNALLF 
      SICGAAFSY LFIMIQMDK DLNKSLRNV TNSIL
    CDS sequence
    • >TC030232 RA
      atgcaagcg ctcactctg acgtcgatt tttgcaccc 
      caacatgtc gtttcaaaa attttgggt gtactaccc 
      ttcacaata caggacggt cgacctgtg ttttccaaa 
      cttggtctt tgtcacagt tttctgtgc atcattttg 
      tacgttttt gtggcccaa agctactac cagacaata 
      tttgatgat acgacctca ggttttatt acattaaaa 
      attttgaaa tttattaca atcgcccgc tatgttggt 
      tacacggcc gtgatgatt aacacattc gttgctgtt 
      ggtctgact tacaaaaaa tttgtcgct ttgttgtcg 
      aaattgtat ggaattgat cgaggattg gggaagttg 
      ggtctagag acatttgtt gtagcaagt ggttacagg 
      attaagaaa aaaatgttc ttgttggtg attttggag 
      tatttaatt ggaaatata gtgtcgatt ttttcatcg 
      caattcaac gacatttcc ggtgctttt gttttactt 
      taccctcga atgattggt tgtaacttg agtgtgacc 
      tattttggt accactttg ttactggaa gagagattc 
      aaattggtc aataagttc ctacttggg gagtgtttc 
      aagtcacga aattgtacc aaatatcca accgatcca 
      aatttttgc gacaaaatc aaattatcg gtcaatatt 
      cacaaagaa ctgaccaga atttgtaga gctatcaac 
      tcgattttt tcagttttt tttctgctt tgggtcact 
      gtgaatttc ttgtgcttg gtttcagat ctgtacatt 
      gttgcttat ttaatctca agcggccag ttttttaat 
      tattggcaa ggcgagttg aaagctttt aagaacgtt 
      tgtatttat tcgattgag atgttttat attgcgaga 
      agatcgtca cggctttgt tctcaagca aacagaact 
      aaaactatt ttggttggg ttgaaagtg gatttaagc 
      aaagaggag gaacgaaat actattatt acttcgagt 
      cttgaactt cgacagttt actcttcaa ataacggct 
      tgtagactt tttacgatt gataatgcg ttacttttt 
      tcgatctgt ggagctgcc ttttcatac ctcttcata 
      atgatacaa atggacaaa gacttgaat aagtctctc 
      cgaaatgta acaaattct attctatag
    mRNA sequence
    • >TC030232 RA
      gacaaatac aatcaataa cccattatt ataattttc 
      aaatggttg aatttggtt tttaatgcc agttcaact 
      gaagcaatg caagcgctc actctgacg tcgattttt 
      gcaccccaa catgtcgtt tcaaaaatt ttgggtgta 
      ctacccttc acaatacag gacggtcga cctgtgttt 
      tccaaactt ggtctttgt cacagtttt ctgtgcatc 
      attttgtac gtttttgtg gcccaaagc tactaccag 
      acaatattt gatgatacg acctcaggt tttattaca 
      ttaaaaatt ttgaaattt attacaatc gcccgctat 
      gttggttac acggccgtg atgattaac acattcgtt 
      gctgttggt ctgacttac aaaaaattt gtcgctttg 
      ttgtcgaaa ttgtatgga attgatcga ggattgggg 
      aagttgggt ctagagaca tttgttgta gcaagtggt 
      tacaggatt aagaaaaaa atgttcttg ttggtgatt 
      ttggagtat ttaattgga aatatagtg tcgattttt 
      tcatcgcaa ttcaacgac atttccggt gcttttgtt 
      ttactttac cctcgaatg attggttgt aacttgagt 
      gtgacctat tttggtacc actttgtta ctggaagag 
      agattcaaa ttggtcaat aagttccta cttggggag 
      tgtttcaag tcacgaaat tgtaccaaa tatccaacc 
      gatccaaat ttttgcgac aaaatcaaa ttatcggtc 
      aatattcac aaagaactg accagaatt tgtagagct 
      atcaactcg attttttca gtttttttt ctgctttgg 
      gtcactgtg aatttcttg tgcttggtt tcagatctg 
      tacattgtt gcttattta atctcaagc ggccagttt 
      tttaattat tggcaaggc gagttgaaa gcttttaag 
      aacgtttgt atttattcg attgagatg ttttatatt 
      gcgagaaga tcgtcacgg ctttgttct caagcaaac 
      agaactaaa actattttg gttgggttg aaagtggat 
      ttaagcaaa gaggaggaa cgaaatact attattact 
      tcgagtctt gaacttcga cagtttact cttcaaata 
      acggcttgt agacttttt acgattgat aatgcgtta 
      cttttttcg atctgtgga gctgccttt tcatacctc 
      ttcataatg atacaaatg gacaaagac ttgaataag 
      tctctccga aatgtaaca aattctatt ctatagttt 
      gcgaaataa ataaacaat ctcatgcaa c
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY