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Gene information
General information
TC000528 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_007417.3: 10150533..10152299
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
    TC000528
    iBeetle screen iB_09648

    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 with embryonic tissue potentially present, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 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): 50% - 80%
    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 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
    • >TC000528 RA
      MDDNLDDYD FTPSSSSNL ASLFGNNSP HSETASLIY 
      TAPKQPKAG GSSEVKNEA PKAESSVVC AKVVQVWKL 
      VEGKYQALG KYGLAVIGS TVQNSYEII LYKEKRNVI 
      MRAKVSSSF EVNLKKDDF ASFYDNIQE NWLIKFNNN 
      DDRMAVMNV IEKFGCVVN RVVEKQDLK TEKTDEVVT 
      PENKAKADI LSRIAKMGQ SILPNKDLD TDNDSVESA 
      DSTVSTHQV SSTSAVIKT SHQIQNVPN NFVMSQPMV 
      YDPLNLLFA ENRAQNSEV RLNLSQISD KLNQIISLV 
      DSDKPSDNS NSLKSRLKV LELRNENLL RELGSSQEE 
      NAKLKLQLK EHESNLKSE EENKLELRL LEEQETVKR 
      EKLEKEICA HKDEIAALS KTVEKQIEE IEACKQSEL 
      AQKVEISRL ENTISDLNT KLQGYENKE ARPEICEEK 
      NAKFIASLK DAMNNMYAN IMSSCSEDQ QTEIGKIVA 
      QNIKTTTLQ IIQNFQQEY ES
    CDS sequence
    • >TC000528 RA
      atggatgat aacttagac gactacgac tttacccct 
      tcgtctagt tcaaatttg gcttcatta tttggcaat 
      aattcacca cattccgaa accgctagc ctcatatac 
      actgccccc aaacaacca aaggctgga ggtagttcc 
      gaagtaaaa aacgaggcc ccaaaggca gaaagcagt 
      gttgtttgt gcaaaagtc gtccaagtt tggaaatta 
      gttgagggt aaataccaa gcgttgggc aaatatggc 
      cttgctgtt attggatca acagttcaa aattcctat 
      gaaattatt ttgtataaa gaaaagcgt aatgtgata 
      atgagggct aaagtatcg tctagtttt gaagtgaac 
      ttaaagaag gacgatttt gcttcgttt tacgacaat 
      attcaagag aactggttg atcaaattt aacaataat 
      gatgaccgt atggctgtt atgaacgtt attgaaaag 
      ttcgggtgt gttgtaaat cgtgtagtt gaaaaacaa 
      gacctcaaa actgaaaaa actgatgaa gttgtaacg 
      ccggaaaat aaagctaag gctgatata ttgtcaaga 
      attgccaaa atggggcag tctatttta ccgaataaa 
      gacttagat acagataat gattctgtt gaaagtgca 
      gacagtaca gtatccacc caccaagtc agctcaaca 
      tcagctgtt atcaagact tcacatcaa attcaaaac 
      gttccaaat aattttgtt atgagtcaa ccgatggtt 
      tacgaccct ttaaatttg ttgtttgct gaaaaccgt 
      gcacaaaat agtgaagtc aggttaaat ttatcgcaa 
      atttcggac aaattaaat cagattatt agtttggtt 
      gactcagat aagcccagt gacaactca aatagcttg 
      aaaagtaga ctcaaagtg ttggagttg cgtaatgaa 
      aacttgttg agggaattg ggatcaagt caggaagaa 
      aacgcaaaa ctcaaatta caattgaag gaacacgaa 
      agtaatttg aagagtgag gaagagaat aaactggaa 
      ttgaggctt ttggaggaa caagaaacc gtcaaacga 
      gagaaattg gaaaaagaa atttgtgca cacaaggac 
      gaaatcgct gccctttct aaaaccgtg gagaaacaa 
      atagaagaa attgaggct tgcaagcag tcagaatta 
      gctcagaaa gttgaaatt tcacgcctc gaaaatacc 
      atttctgat ttgaacaca aaattgcaa ggttatgag 
      aataaggaa gcaagacca gaaatttgt gaggaaaaa 
      aatgcgaaa tttattgcg tcactcaag gatgcgatg 
      aataatatg tacgcaaat attatgtca agttgcagc 
      gaagatcag caaaccgaa atcggcaaa attgttgcc 
      caaaacatc aaaaccacg acactacaa ataattcaa 
      aatttccaa caagagtat gaaagttag
    mRNA sequence
    • >TC000528 RA
      caataaaat gtcaaattg tttgacaat taaaaaagt 
      caggttaag actaaccct ttattttta tttcgtaat 
      tgttcaggt ttttttaat ttaatctat gtataaaat 
      gtgttaaaa atggatgat aacttagac gactacgac 
      tttacccct tcgtctagt tcaaatttg gcttcatta 
      tttggcaat aattcacca cattccgaa accgctagc 
      ctcatatac actgccccc aaacaacca aaggctgga 
      ggtagttcc gaagtaaaa aacgaggcc ccaaaggca 
      gaaagcagt gttgtttgt gcaaaagtc gtccaagtt 
      tggaaatta gttgagggt aaataccaa gcgttgggc 
      aaatatggc cttgctgtt attggatca acagttcaa 
      aattcctat gaaattatt ttgtataaa gaaaagcgt 
      aatgtgata atgagggct aaagtatcg tctagtttt 
      gaagtgaac ttaaagaag gacgatttt gcttcgttt 
      tacgacaat attcaagag aactggttg atcaaattt 
      aacaataat gatgaccgt atggctgtt atgaacgtt 
      attgaaaag ttcgggtgt gttgtaaat cgtgtagtt 
      gaaaaacaa gacctcaaa actgaaaaa actgatgaa 
      gttgtaacg ccggaaaat aaagctaag gctgatata 
      ttgtcaaga attgccaaa atggggcag tctatttta 
      ccgaataaa gacttagat acagataat gattctgtt 
      gaaagtgca gacagtaca gtatccacc caccaagtc 
      agctcaaca tcagctgtt atcaagact tcacatcaa 
      attcaaaac gttccaaat aattttgtt atgagtcaa 
      ccgatggtt tacgaccct ttaaatttg ttgtttgct 
      gaaaaccgt gcacaaaat agtgaagtc aggttaaat 
      ttatcgcaa atttcggac aaattaaat cagattatt 
      agtttggtt gactcagat aagcccagt gacaactca 
      aatagcttg aaaagtaga ctcaaagtg ttggagttg 
      cgtaatgaa aacttgttg agggaattg ggatcaagt 
      caggaagaa aacgcaaaa ctcaaatta caattgaag 
      gaacacgaa agtaatttg aagagtgag gaagagaat 
      aaactggaa ttgaggctt ttggaggaa caagaaacc 
      gtcaaacga gagaaattg gaaaaagaa atttgtgca 
      cacaaggac gaaatcgct gccctttct aaaaccgtg 
      gagaaacaa atagaagaa attgaggct tgcaagcag 
      tcagaatta gctcagaaa gttgaaatt tcacgcctc 
      gaaaatacc atttctgat ttgaacaca aaattgcaa 
      ggttatgag aataaggaa gcaagacca gaaatttgt 
      gaggaaaaa aatgcgaaa tttattgcg tcactcaag 
      gatgcgatg aataatatg tacgcaaat attatgtca 
      agttgcagc gaagatcag caaaccgaa atcggcaaa 
      attgttgcc caaaacatc aaaaccacg acactacaa 
      ataattcaa aatttccaa caagagtat gaaagttag 
      actgaggga aaataatca tttccaatt atttgtgat 
      acgtattat attcgttaa aaaatacta caaaataaa 
      caattcaaa tgcat
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY