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Gene information
General information
TC003524 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_seqid16: 1676745..1678589
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
    TC003524
    iBeetle screen iB_00580
    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)

    pupa eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (3 dpi)
    Days post injection


    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 50
    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: 20.0% (includes death as larva, prepupa, pupa, adult)

    adult lethality - Number of animals showing the phenotype: 1 (16 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 1 (19 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC003524 RA
      MALRNGVRR GGALISNYF RSYSSAGPL VNVAVNEKT 
      GVATATLNR PPVNSMNLE LLTEISNTL TQLEQNKCR 
      GLILTSSSK SVFSAGLDI LEMYKPDPS RLKEWWTAL 
      QQTWIKLYG SSYPTVAVI NGHAPAGGC LLSLSCEYR 
      IMLNNFTIG LNETQLGIV APQWFIASM RNVIGTRQT 
      ELALTSGNL FSTDEALKI GMIDEVAAS KEDGIARAE 
      KFCGRFARI PPIARARSK LGVRGADIQ DLTNNLQKD 
      YENVANYVA QDSVQKGLE MYLQSLKQK QK
    CDS sequence
    • >TC003524 RA
      atggctctg agaaacggt gttcgtcgc ggtggtgcc 
      ttaatttct aattatttc cgaagttac tcaagcgct 
      gggcccctt gttaacgtc gccgttaac gaaaaaaca 
      ggggttgcg acggcaacc ctcaaccgc ccccctgtg 
      aacagcatg aatttggag ctccttact gaaatatca 
      aacactttg actcaactg gaacagaac aagtgtaga 
      gggttaatt ttgacctcg agttccaag agtgtcttc 
      agtgccggc ttggacatc cttgagatg tacaagccg 
      gaccccagc cgcttgaaa gaatggtgg actgcttta 
      cagcaaact tggattaaa ctttacggt tcttcgtac 
      cctactgtg gcagttatc aatggccac gccccagct 
      gggggctgt ttgctgtcc ttgtcgtgt gaatacaga 
      ataatgttg aacaatttc accattggc ttaaacgag 
      acccaactt gggattgtg gccccacaa tggttcatt 
      gcgtccatg aggaatgtg attgggacg agacagact 
      gaattggct ttaactagc ggcaatctt ttctcaact 
      gacgaagct ttgaaaatt ggaatgatt gacgaggtg 
      gcggctagc aaagaggat ggcattgca agagcggag 
      aagttttgc ggaagattc gccagaatt ccgccaatt 
      gcaagagcc agatccaag ttgggagtg cgaggcgct 
      gacatacaa gacctcaca aacaatctc caaaaagat 
      tacgaaaat gtggcgaac tacgtagct caggacagt 
      gtgcaaaag gggttggaa atgtatctg caatcattg 
      aagcagaaa cagaaataa
    mRNA sequence
    • >TC003524 RA
      caacgcaac gtaagtcgt tgatcaaaa tgacaagct 
      gggtataaa ggttatgtt cacataacc agaagttat 
      tttttaatt tagtagttt tattgtggt gtcgttacc 
      taattcctt tcccggata ttgttttaa ttttattcc 
      cctgcaaaa tggctctga gaaacggtg ttcgtcgcg 
      gtggtgcct taatttcta attatttcc gaagttact 
      caagcgctg ggccccttg ttaacgtcg ccgttaacg 
      aaaaaacag gggttgcga cggcaaccc tcaaccgcc 
      cccctgtga acagcatga atttggagc tccttactg 
      aaatatcaa acactttga ctcaactgg aacagaaca 
      agtgtagag ggttaattt tgacctcga gttccaaga 
      gtgtcttca gtgccggct tggacatcc ttgagatgt 
      acaagccgg accccagcc gcttgaaag aatggtgga 
      ctgctttac agcaaactt ggattaaac tttacggtt 
      cttcgtacc ctactgtgg cagttatca atggccacg 
      ccccagctg ggggctgtt tgctgtcct tgtcgtgtg 
      aatacagaa taatgttga acaatttca ccattggct 
      taaacgaga cccaacttg ggattgtgg ccccacaat 
      ggttcattg cgtccatga ggaatgtga ttgggacga 
      gacagactg aattggctt taactagcg gcaatcttt 
      tctcaactg acgaagctt tgaaaattg gaatgattg 
      acgaggtgg cggctagca aagaggatg gcattgcaa 
      gagcggaga agttttgcg gaagattcg ccagaattc 
      cgccaattg caagagcca gatccaagt tgggagtgc 
      gaggcgctg acatacaag acctcacaa acaatctcc 
      aaaaagatt acgaaaatg tggcgaact acgtagctc 
      aggacagtg tgcaaaagg ggttggaaa tgtatctgc 
      aatcattga agcagaaac agaaataag gatattata 
      agttgatat tcttttata taatgaagt gtaaaattt 
      attatccaa accaagaat atgtgt