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Gene information
General information
TC000475 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: 5329257..5330384
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
    TC000475
    iBeetle screen iB_00104
    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)

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


    adult lethality - Number of animals showing the phenotype: 1 (9 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: 10.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 10.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC000475 RA
      MEDANIQQG KPFKITDSS RETRKGVVA 
      GSLEDLTSK VIEKLGISG DITVVLEAD 
      GTEIDDEEY FATLEPHTS LMILNGGQK 
      WMPPAPPCR LSTDQPDDG KGGAELAGL 
      VGKLKHNLC HVSLLGGAE LELLSDMDP 
      DSLVDITFP DKIFLEQLK EASGRFLSE 
      KRQAQDAMD LLRLYQEKE AESGETN
    CDS sequence
    • >TC000475 RA
      atggaagat gcaaacatc cagcaaggc aagcctttc 
      aaaatcacc gactcctcc cgcgagacc aggaagggg 
      gtcgttgcc ggcagcctc gaggaccta acctccaaa 
      gtcatcgaa aagctcggc atttctggt gatattacc 
      gtcgtcctg gaagcggac ggcaccgaa atcgacgac 
      gaggagtat ttcgctacg cttgagccg cacaccagt 
      cttatgatc ctgaacggg ggccagaag tggatgccc 
      ccggcgccc ccttgccgc ctctccacc gaccagccc 
      gacgacggc aagggcggc gccgagctg gccggcctc 
      gtcggcaaa ctcaagcac aatttgtgc cacgtgtcg 
      ctcttaggg ggcgccgag ttggagctg ttgagtgac 
      atggacccg gactcgcta gttgacatc accttcccg 
      gacaagatc ttcctcgag cagctcaag gaggcgtcg 
      gggaggttc ttgagcgag aagcggcaa gcacaagat 
      gccatggat ttgctaagg ctgtaccag gagaaagag 
      gctgagtcc ggggagacc aactga
    mRNA sequence
    • >TC000475 RA
      cattagggc ggcaaccac atttcaaac aaagaattc 
      gtgcagcac acacgactc actttttaa atgtaaacc 
      ttggtaaat acaatggaa gatgcaaac atccagcaa 
      ggcaagcct ttcaaaatc accgactcc tcccgcgag 
      accaggaag ggggtcgtt gccggcagc ctcgaggac 
      ctaacctcc aaagtcatc gaaaagctc ggcatttct 
      ggtgatatt accgtcgtc ctggaagcg gacggcacc 
      gaaatcgac gacgaggag tatttcgct acgcttgag 
      ccgcacacc agtcttatg atcctgaac gggggccag 
      aagtggatg cccccggcg cccccttgc cgcctctcc 
      accgaccag cccgacgac ggcaagggc ggcgccgag 
      ctggccggc ctcgtcggc aaactcaag cacaatttg 
      tgccacgtg tcgctctta gggggcgcc gagttggag 
      ctgttgagt gacatggac ccggactcg ctagttgac 
      atcaccttc ccggacaag atcttcctc gagcagctc 
      aaggaggcg tcggggagg ttcttgagc gagaagcgg 
      caagcacaa gatgccatg gatttgcta aggctgtac 
      caggagaaa gaggctgag tccggggag accaactga 
      tccacgctt aacgacgca ttggaaatc ccgttttaa 
      aaatgtaag tgcttttac ggagatgtg gagtgtttt 
      ttttttggt gttttgtag ggctgcctc acaccccga 
      gtatttaaa agactgaat tgccaataa attactgtc 
      ctggcgtaa cccaattgc tcaatcact gcacggcca 
      cgttcacac taacaccaa gcaacgttg gttgacaca 
      aattctaac atcgcaaca ataggactt tttcaccga 
      gggcacgtc acatatacc cacacattg taatcacca 
      ctccaccta ttattcatg tttgtataa gtctcttat 
      tagtgaatt acgcatgtt accctagct gtgatacaa 
      ctgtagtac tcgactttt caaactagt tcaacccct 
      aatcaaaat tcactaaca agaggagcg tctcattca 
      taacatagt ttttgctat aatatgaat taatataat 
      catatactt gta