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Gene information
General information
TC012328 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_007424.3: 4868059..4869195
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
    TC012328
    iBeetle screen iB_05215

    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: 10.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - 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: 40.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 50.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality - Number of animals showing the phenotype: 4 (11 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC012328 RA
      MAGLFSTIV RRGPQLLKT AIPAGKNHA 
      FRNFSVAPT LCAPRVQHP APDFKGTAV 
      INDGFKEIQ LSDYKGKYV VLVFYPLDF 
      TFVCPTELI ALDERYDDF KNLNAEVIG 
      CSIDSHFSH LGWMNTKRS EGGLGKLRY 
      PLLSDINKT IARDYDVLL EKEGIALRG 
      LFIIDPNGI LRQITVNDL PIGRSVDEA 
      LRLIEAIQF FEKNGEVCP ANWKKGSKT 
      IKPDPQGSK EYFQAANK
    CDS sequence
    • >TC012328 RA
      atggctgga ctatttagc acaattgta cgacggggc 
      ccccaattg ttaaaaact gcgattccg gctggtaaa 
      aatcatgca tttagaaat ttttccgtt gcccccact 
      ttgtgcgct cctcgagtg caacacccc gctcctgac 
      tttaaagga acagcagtt ataaatgat ggctttaaa 
      gagatacaa ttgagcgac tataaggga aaatatgtt 
      gttctagtt ttctatcct cttgatttc acttttgtg 
      tgccccact gaattgata gcgttagat gaacgttac 
      gacgacttc aaaaacttg aacgctgaa gtaattggt 
      tgttctatt gattcacat ttctcgcac ttagggtgg 
      atgaacaca aaaagatct gaagggggt ttgggcaaa 
      ctaagatat cccttgctt tctgacatt aataaaact 
      atagccaga gactatgat gtcctgctc gagaaagag 
      ggaattgcc ttgagaggt ctcttcatt attgaccct 
      aatggcatc cttaggcaa attacggta aatgatttg 
      ccaataggt cgttcagta gatgaggct cttcggtta 
      atcgaagcc attcaattc tttgaaaaa aatggagaa 
      gtgtgtcca gctaattgg aagaaaggt agtaagaca 
      atcaaacct gaccctcaa ggatctaaa gaatatttc 
      caagccgct aataagtaa
    mRNA sequence
    • >TC012328 RA
      gccggcgct caaagtccc tagacttgg cactgactt 
      tccaaagtt tgataatta atttcagcg actcatcgc 
      aattttaaa caaatttgc tggcagtat caacgtttt 
      gttatcatg gctggacta tttagcaca attgtacga 
      cggggcccc caattgtta aaaactgcg attccggct 
      ggtaaaaat catgcattt agaaatttt tccgttgcc 
      cccactttg tgcgctcct cgagtgcaa caccccgct 
      cctgacttt aaaggaaca gcagttata aatgatggc 
      tttaaagag atacaattg agcgactat aagggaaaa 
      tatgttgtt ctagttttc tatcctctt gatttcact 
      tttgtgtgc cccactgaa ttgatagcg ttagatgaa 
      cgttacgac gacttcaaa aacttgaac gctgaagta 
      attggttgt tctattgat tcacatttc tcgcactta 
      gggtggatg aacacaaaa agatctgaa gggggtttg 
      ggcaaacta agatatccc ttgctttct gacattaat 
      aaaactata gccagagac tatgatgtc ctgctcgag 
      aaagaggga attgccttg agaggtctc ttcattatt 
      gaccctaat ggcatcctt aggcaaatt acggtaaat 
      gatttgcca ataggtcgt tcagtagat gaggctctt 
      cggttaatc gaagccatt caattcttt gaaaaaaat 
      ggagaagtg tgtccagct aattggaag aaaggtagt 
      aagacaatc aaacctgac cctcaagga tctaaagaa 
      tatttccaa gccgctaat aagtaatga gtaaacgtt 
      ttgtgtttg taaattgtt acgttgtaa agattttga 
      ataaatata aacttaagt a
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY