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Gene information
General information
TC000520 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.

scaffold_seqid7: 4729011..4737626
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
    TC000520
    iBeetle screen iB_00116
    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)

    pupa eclosion not fulfilled (died as pupa) - 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, no cuticle, but muscle fluorescence - Penetrance (percentage of animals/eggs on slide): less than 30%
    larval musculature fluorescence decreased - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - 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
    egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    legs segment fused - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    legs irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - 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: 0.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 0.0% (includes death as larva, prepupa, pupa, adult)

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC000520 RA
      MNGNVIIVL MSLMSPIAA FTDYYNRPT 
      ARPRARPHI SDTSNRNQY SEATNETIA 
      HMHLDSRSS NSRQQGEDT PIVYPEKPT 
      CDHGLCYNV AGYPKEQIK KILSRSHDM 
      NRYFETSEE VVNLDNRFN TDEMSLCET 
      TIRTIYPEK ANNTQKVEK VIVNVEGHK 
      QGIVFETCV NNGKCKFSS NFPTGYTSY 
      CKQKYIHKR LMVLGDDDK FVFDLFEVP 
      SCCICTVSK TN
    CDS sequence
    • >TC000520 RA
      atgaatggc aacgtgata atagtttta atgtccttg 
      atgtccccc attgctgct ttcacagac tattacaac 
      agacctacc gcaagacca cgagcaaga ccacacatc 
      agtgataca tctaacaga aatcaatat tcagaagct 
      accaatgaa acaatcgcg catatgcat ttagactca 
      agaagttca aacagccgc caacaaggc gaagacact 
      cccattgtt tatcctgaa aagccgact tgcgaccac 
      ggcctgtgc tacaacgtt gctggctat ccaaaagaa 
      caaataaaa aaaatccta agtagatct cacgacatg 
      aaccgatat tttgagact agtgaagaa gtcgtaaac 
      ttagacaac cgatttaat acagacgaa atgtcactg 
      tgtgaaact acaattcgg acaatctat cctgagaaa 
      gcgaacaat actcagaaa gtggagaag gtgattgtc 
      aatgtggag ggacataag caaggaatt gtatttgaa 
      acgtgcgtg aataatgga aaatgtaaa ttcagcagc 
      aattttccg acagggtac acctcctac tgtaagcag 
      aagtacatc cacaagaga ttgatggtt ttaggggac 
      gatgacaaa ttcgtgttt gatctgttc gaagttccg 
      tcgtgttgt atttgtaca gttagcaaa actaattag
    mRNA sequence
    • >TC000520 RA
      acattaccg tttgacttg gtgtccttg tcgatttac 
      aacaaattt taatgtact ctcgaacag cggtagtgt 
      tgaatgttg tggtcggtc agacgctcc gctccggtt 
      aaaacagac gttgggtct ctttagccg ctttcgtat 
      catgcagac ggggtggta gttgctttg tagtgctta 
      gtagtttgg taatagcaa tgaatggca acgtgataa 
      tagttttaa tgtccttga tgtccccca ttgctgctt 
      tcacagact attacaaca gacctaccg caagaccac 
      gagcaagac cacacatca gtgatacat ctaacagaa 
      atcaatatt cagaagcta ccaatgaaa caatcgcgc 
      atatgcatt tagactcaa gaagttcaa acagccgcc 
      aacaaggcg aagacactc ccattgttt atcctgaaa 
      agccgactt gcgaccacg gcctgtgct acaacgttg 
      ctggctatc caaaagaac aaataaaaa aaatcctaa 
      gtagatctc acgacatga accgatatt ttgagacta 
      gtgaagaag tcgtaaact tagacaacc gatttaata 
      cagacgaaa tgtcactgt gtgaaacta caattcgga 
      caatctatc ctgagaaag cgaacaata ctcagaaag 
      tggagaagg tgattgtca atgtggagg gacataagc 
      aaggaattg tatttgaaa cgtgcgtga ataatggaa 
      aatgtaaat tcagcagca attttccga cagggtaca 
      cctcctact gtaagcaga agtacatcc acaagagat 
      tgatggttt taggggacg atgacaaat tcgtgtttg 
      atctgttcg aagttccgt cgtgttgta tttgtacag 
      ttagcaaaa ctaattaga tatcatatt tatgcaagt 
      atatgtact tagtgtagc taatagctg agctgtgta 
      tttatatta aattgccgt ttagcatcg ctattgagt 
      ttcctttaa attagatta ttgtgataa aatgttttg 
      tgtaattaa atcaagaca cttcaatat tgcagccag 
      ttaattaat ttatcccta actgctgat caaaagcaa 
      taaaccacg agacaaggc