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Gene information
General information
TC004565 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_seqid6: 259877..288684
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
    TC004565
    iBeetle screen iB_03789
    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)

    adult lethality - Number of animals showing the phenotype: 2 (9 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 embryo/egg not developed/not fertilized - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    embryo/egg with embryonic tissue potentially present, no cuticle, no fluorescence - 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): more than 80%
    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%
    abdomen posterior remaining - 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%
    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: 30.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 30.0% (includes death as larva, prepupa, pupa, adult)

    pupal molt not fulfilled (died as L7 or prepupa) - Number of animals showing the phenotype: 3 (11 dpi)
    Days post injection


    Fertility
    fertility after larval injection decreased (no living offspring) -
    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC004565 RA
      MDADNYTQN VQNDVPPEV WEIIHEFLL GQANPKLDL 
      RKPHLRPSV ASSYPFILF MYSLLIVTG VALNVSVFI 
      HIVRNKLQN DVTCSFVLN NVISDIVKC VVVLPITLY 
      VLLINNWIL GELLCSFLP MLQDIPMHT TTLTFLLMA 
      WDRLRFIRH PTKARFPAF VCIIGTWLA SVCLVLPYP 
      IYIIYIDLG KYVPSLDGV GICLVNLAD DMQEYMRGI 
      FLIMYAAPL ATISYLYVR VSRELQTQQ SSLTVIKFE 
      TRREGRSRT DSHSTTTDF RSSRGDYDR SSQYSRSSY 
      RDHENGRTR YENEEPDLD VAKEKRTQK YLVPMVTLY 
      AIYLCPLMI LRLARLVLP ETYENNKNF DISYTMFVW 
      VAFAPTCST PVLYIVWQM NRSSKERLL GYFRFSNRK 
      LRQSCETVI TAAGTPAVP RAKCGQTDT IFLRPDDIK 
      GGITQWGIP VSSEWNN
    CDS sequence
    • >TC004565 RA
      atggacgcg gacaactac acccaaaac gtccaaaac 
      gacgtcccg cccgaagtt tgggaaatc atccacgag 
      tttttgctc ggccaagcg aacccgaaa ctagatctg 
      cggaaacca catcttaga ccttctgta gcgagcagt 
      taccctttc atattgttc atgtactcc cttttgatc 
      gtcacggga gtggcgctc aacgtttcg gttttcatc 
      cacatcgta agaaataaa cttcaaaac gatgtcact 
      tgtagtttc gtcctcaac aatgtcatt agtgatatt 
      gtcaaatgt gtggtagtt ttgcccata acgctctac 
      gttctgttg attaataac tggattctg ggcgaactg 
      ctctgctcg tttctgccc atgttgcag gacattcca 
      atgcataca acgactcta acgttcctg ttgatggca 
      tgggacaga ctccggttt attagacac ccaacgaaa 
      gcacgtttt ccagctttt gtctgtatt attggaaca 
      tggctagcg tctgtgtgc ttagtgctg ccctatccc 
      atctacatt atttatatc gatttgggg aaatacgtc 
      ccatcattg gatggtgtg ggaatttgt ctcgtcaac 
      ttggccgac gacatgcag gaatatatg cgaggaatt 
      ttcctgata atgtatgcg gctcctctc gccactatt 
      tcttactta tacgtaaga gtttcccgc gaattgcag 
      actcagcag agttccctg acagtgata aaatttgaa 
      actcggcga gaaggacgt tccaggacc gattcgcac 
      tcgacgaca acggatttc cgaagcagc cgtggcgat 
      tacgaccgt tcgagtcaa tacagcaga agtagttat 
      cgcgatcac gaaaacggc cgcacacgg tacgaaaac 
      gaagagccc gacttggat gtggctaaa gagaaacgc 
      actcagaag tacttagtc ccgatggtt accctttac 
      gctatctac ttgtgtcct ctaatgatc ctccgcttg 
      gcccgtctc gtccttccc gagacgtac gaaaataat 
      aaaaacttt gacatttcg tacacaatg ttcgtgtgg 
      gtggcgttc gcacccacc tgttccact cctgtcctt 
      tacatcgtt tggcagatg aacagatct tcaaaggag 
      cgtttgttg ggatacttc cgattcagc aatcggaag 
      ctgagacag tcgtgcgaa acggtaata acagctgcc 
      gggacccca gccgtgcca agagcgaaa tgcggtcag 
      actgacacg attttcctt cgtcctgat gatataaag 
      ggaggcatc actcaatgg ggaattcct gtctcttcc 
      gagtggaac aattga
    mRNA sequence
    • >TC004565 RA
      aaacttgtc gcttgtgct tggtgaaag tgatcgcaa 
      aatgtgaaa acaattttt tctggcttt gaaatctgg 
      cggagaaaa ttatggacg cggacaact acacccaaa 
      acgtccaaa acgacgtcc cgcccgaag tttgggaaa 
      tcatccacg agtttttgc tcggccaag cgaacccga 
      aactagatc tgcggaaac cacatctta gaccttctg 
      tagcgagca gttaccctt tcatattgt tcatgtact 
      cccttttga tcgtcacgg gagtggcgc tcaacgttt 
      cggttttca tccacatcg taagaaata aacttcaaa 
      acgatgtca cttgtagtt tcgtcctca acaatgtca 
      ttagtgata ttgtcaaat gtgtggtag ttttgccca 
      taacgctct acgttctgt tgattaata actggattc 
      tgggcgaac tgctctgct cgtttctgc ccatgttgc 
      aggacattc caatgcata caacgactc taacgttcc 
      tgttgatgg catgggaca gactccggt ttattagac 
      acccaacga aagcacgtt ttccagctt ttgtctgta 
      ttattggaa catggctag cgtctgtgt gcttagtgc 
      tgccctatc ccatctaca ttatttata tcgatttgg 
      ggaaatacg tcccatcat tggatggtg tgggaattt 
      gtctcgtca acttggccg acgacatgc aggaatata 
      tgcgaggaa ttttcctga taatgtatg cggctcctc 
      tcgccacta tttcttact tatacgtaa gagtttccc 
      gcgaattgc agactcagc agagttccc tgacagtga 
      taaaatttg aaactcggc gagaaggac gttccagga 
      ccgattcgc actcgacga caacggatt tccgaagca 
      gccgtggcg attacgacc gttcgagtc aatacagca 
      gaagtagtt atcgcgatc acgaaaacg gccgcacac 
      ggtacgaaa acgaagagc ccgacttgg atgtggcta 
      aagagaaac gcactcaga agtacttag tcccgatgg 
      ttacccttt acgctatct acttgtgtc ctctaatga 
      tcctccgct tggcccgtc tcgtccttc ccgagacgt 
      acgaaaata ataaaaact ttgacattt cgtacacaa 
      tgttcgtgt gggtggcgt tcgcaccca cctgttcca 
      ctcctgtcc tttacatcg tttggcaga tgaacagat 
      cttcaaagg agcgtttgt tgggatact tccgattca 
      gcaatcgga agctgagac agtcgtgcg aaacggtaa 
      taacagctg ccgggaccc cagccgtgc caagagcga 
      aatgcggtc agactgaca cgattttcc ttcgtcctg 
      atgatataa agggaggca tcactcaat ggggaattc 
      ctgtctctt ccgagtgga acaattgat ttgaatcgg 
      ctccatcgc acgtttttg tggagagcc gttcattgt 
      actcgatgt aaaatgtca gcgtgatcg gtctgactg 
      taataatgc gaaatatcc agcgagttt cgatactgt 
      gtctatttt tacttgttg tacgtcaca atttacagt 
      tggaaaaaa attaattgt gaagagcat cacttcaaa 
      aaacgaact gcaacattt ttccctcga tgataaatt 
      tcaagggca gattttacg acgcaggtt aatgtcaga 
      tttaagtat tttaaggcc acctggaga tttattctt 
      tttttaatg tctcctttg gcaaaagct ttccggtgg 
      aaagtatga gaagggtcg agggttgtt tgtggtcgg 
      aatttaaac ccttaaaac