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Gene information
General information
TC008221 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_seqid33: 6951150..6952459
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
    TC008221
    iBeetle screen iB_01320
    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 muscle fluorescence/pattern, but not hatched (pot. late embryonic lethality) - Penetrance (percentage of animals/eggs on slide): less than 30%

    embryo/egg not developed/not fertilized - Penetrance (percentage of animals/eggs on slide): less than 30%
    embryo/egg with embryonic tissue, no cuticle, but muscle fluorescence - Penetrance (percentage of animals/eggs on slide): less than 30%

    embryo/egg with embryonic tissue, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): less than 30%

    larval musculature pattern irregular - 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): more than 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): more than 80%
    head anterior not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 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: 10.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 20.0% (includes death as larva, prepupa, pupa, adult)

    pupal molt delayed (still larva) - Number of animals showing the phenotype: 3 (11 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 1 (22 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 1 (41 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC008221 RA
      MAESEAKSG GFLTNIVLE IVQSPVNIL 
      LVCLIGILV YKIVKSRQD VPQSRQPEP 
      ELPKLKKRD FTVEELKKY DGNQEDGRV 
      LVAVNGNVY DVTKGKRFY GPGGPYAAF 
      GGRDASRGL ATFSVSAKT DEYDDLSDL 
      NSMEMDSVR EWEAQFKEK YDLVGKLLK 
      PGEQPTNYS DDEEDETPT GAGDKSKDD
    CDS sequence
    • >TC008221 RA
      atggcggaa agcgaagca aaatccggc ggttttctt 
      acaaatatt gtcttggaa atcgtccaa agtcccgta 
      aacattctt ttagtgtgt ctgatcgga attctagtg 
      tacaaaata gtgaaaagc cgacaagat gtgcctcag 
      agtcgccaa cccgagccg gagctcccc aaattgaaa 
      aaaagagac tttaccgtt gaagaactg aaaaaatac 
      gacggaaac caagaggat ggacgcgtc ctcgtggcc 
      gtcaacggc aacgtctac gacgttaca aaagggaag 
      cgtttttac ggccccggg ggcccttat gcggcgttc 
      ggcggccgc gacgcctcc agggggctc gccaccttc 
      tcagtctcg gcaaaaacc gacgagtac gacgatttg 
      agcgacctt aacagcatg gaaatggat tcggtccgg 
      gaatgggag gcgcagttc aaagagaaa tacgacctc 
      gttgggaag ctactgaag ccaggcgag cagcccacg 
      aactactct gatgatgaa gaagatgag accccaacg 
      ggcgcgggc gataaaagc aaagatgat tga
    mRNA sequence
    • >TC008221 RA
      acaccggtg aaaaaatga ctatgaagt tgtcaaatc 
      aagtcagat agttggtca agatggcgg aaagcgaag 
      caaaatccg gcggttttc ttacaaata ttgtcttgg 
      aaatcgtcc aaagtcccg taaacattc ttttagtgt 
      gtctgatcg gaattctag tgtacaaaa tagtgaaaa 
      gccgacaag atgtgcctc agagtcgcc aacccgagc 
      cggagctcc ccaaattga aaaaaagag actttaccg 
      ttgaagaac tgaaaaaat acgacggaa accaagagg 
      atggacgcg tcctcgtgg ccgtcaacg gcaacgtct 
      acgacgtta caaaaggga agcgttttt acggccccg 
      ggggccctt atgcggcgt tcggcggcc gcgacgcct 
      ccagggggc tcgccacct tctcagtct cggcaaaaa 
      ccgacgagt acgacgatt tgagcgacc ttaacagca 
      tggaaatgg attcggtcc gggaatggg aggcgcagt 
      tcaaagaga aatacgacc tcgttggga agctactga 
      agccaggcg agcagccca cgaactact ctgatgatg 
      aagaagatg agaccccaa cgggcgcgg gcgataaaa 
      gcaaagatg attgaattt gaggctgaa agactgctc 
      ttctagtgt aaaaaaata tcttcgttt tatatgatc 
      ttgttggct tttttaaca tatttttta aggatggtt 
      cgtgtgatg catttccgg taattgtta cttatgtga 
      gcttctttt caaattttt atgtaacgc taaagtggt 
      aacgattcg attccagtt tgtgcattt tctaatttt 
      aagtctcgt cataaatta cggtccctt ttccgccaa 
      taaattcgc aatattgta acaacactg caaatgacg 
      tattttatt tttgtgttg ggtaatttt aagatttag 
      cgagtgcca caagtgggt caaggttac aacacacgt 
      taccaactt cagtgtggc tttagttgg taacatgtg 
      gcacttttt attttaggg gctacagtt tttaatata 
      cggcacgtt tcaaatgtc tattttctt aacgaaaca 
      ccagtactt aggtaacac aggaccgta ctttgtaac 
      atttgaaat gtacctttc ttttatgac cagctacat 
      aattatggt gtgtgtata taattgtat gtatgaaga 
      gtttttgaa gaaattata catgttaac