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Gene information
General information
TC008996 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_007422.5: 9003099..9037729
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
    TC008996
    iBeetle screen iB_08747

    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 embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    labrum not fused - Penetrance (percentage of animals/eggs on slide): more than 80%

    mandible potentially shape irregular - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    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: % (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: % (includes death as larva, prepupa, pupa, adult)

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC008996 RA
      MSAAANYGE GPVVTDRGD LHGLLPELN GVELAMSLS 
      PKHHHQGGP QVLSHLQQL HHSTPFSVT DILSPIEES 
      YRKLELAAN PPSPYRSTS SGSSINNSP GAGGTPGGG 
      GCSMNSSYP VMGQFGGGQ YCPVSENLG LTPHYSSGW 
      YQSSAADPR FAISRLMGS SAASANMGH GLNTGMPGL 
      GACAMPDSK PMQFPLAQR RKRRVLFTQ AQVYELERR 
      FKQQKYLSA PEREHLASL IHLTPTQVK IWFQNHRYK 
      CKRQAKEKA MAEQNQHNQ SSSSPRRVA VPVLVKDGK 
      PCSAGSGST QTSSSATSS DTSRTQQSP VCQPQQQAQ 
      CMGNGSLAM AYRQQNNYQ QQCGTYLPL QTRAW
    CDS sequence
    • >TC008996 RA
      atgtctgca gctgccaac tacggggaa gggcccgtg 
      gtgacggac cggggcgac ctccatggg ttgctgccc 
      gagctcaat ggggtcgag ctggctatg agcttgagt 
      cccaagcat catcatcaa ggtggaccg caagtcttg 
      agccatctc cagcagttg catcattcg acgccgttc 
      agtgttacc gatattttg agtcccatc gaggagtcg 
      tacaggaaa ttggagttg gcggccaat ccgccgtcg 
      ccctacagg tctacatcc agcggaagt agtatcaac 
      aactccccc ggagccggt ggcactccc ggaggcggc 
      ggctgcagc atgaacagc agttatccc gtaatgggg 
      cagttcggc ggtggccaa tactgccca gtgtccgaa 
      aacctaggt ctcaccccc cactattca tcaggatgg 
      taccaaagc tcggcagca gacccccga tttgcaatt 
      tcccgctta atgggcagc tctgcagcc tcggcgaac 
      atgggtcac gggctgaac accggcatg ccgggcctg 
      ggcgcctgc gccatgccc gactccaag cccatgcag 
      ttccccttg gcccaacgg cggaagagg cgggtcctc 
      ttcacgcaa gcacaggtt tacgagctc gagaggagg 
      ttcaagcag cagaagtat ctgtcagcc cccgaaagg 
      gagcatctg gcctcgttg attcatctc acgcctacc 
      caggtgaaa atctggttc cagaaccac cggtacaag 
      tgcaaaaga caagccaaa gaaaaggcg atggccgaa 
      caaaaccaa cataatcag tcctccagc tcgccgcgt 
      cgggtggcc gtgcccgtg ctggtcaaa gacgggaag 
      ccctgctcc gccggctcg gggagcacc cagacctcc 
      tcttcagcg acctcgtcc gacacatct cgcacccaa 
      caatccccc gtctgccag ccccaacaa caggcccaa 
      tgcatgggc aacggcagc ttggccatg gcataccga 
      caacaaaac aactaccaa caacaatgt gggacttat 
      ctgcccctt caaacaagg gcgtggtag
    mRNA sequence
    • >TC008996 RA
      gaccgacgc caccgccgc tctattatc ataatataa 
      tcaaagagc tcgcattac gtcaagtgg ttccgagtg 
      ttgctgttg acaacaatc gtctttgca ctcgttctc 
      ctatgtcct tatcttgct cctcgactt cctcggtag 
      catcggatt tcgtccctg aatcaaaac ggcgagaaa 
      aagtgtgct ccatcccaa atcaatatg cccaataaa 
      acgatataa atacacaat atcaaagtg aagagcggc 
      ccctctcga gtagtcccc gcccctttg ttccgtaaa 
      acctcgccc cataagacc gtttttccc gtgtaaatc 
      gctctgaaa gatcgctag gtaagaggg cgattccca 
      gctggaaac aaacgaagg tgcggcttg gtgcgattg 
      gtcgattcg ggcagtccc cgcccacct tccgcttcg 
      agtacccag aatgcagcg gggcccgac agaagtgcc 
      atgtacact ttgaggaca ctctaagag tacgcttag 
      gcaagttaa gcgcacttc gccgttaaa gagtcttac 
      atcaagagt gctctaaag agtgctata gctcgatca 
      ctgatttat ttatatagt tagtcagtt tgtttattg 
      aattcggat atcggaacg accggaagc cgggcgctt 
      gtacttctt cgtgcataa tcgatgacg ggcgtttag 
      tgcggtggg tgagtgccc ttcagcggt cctggagtc 
      catgtctgc agctgccaa ctacgggga agggcccgt 
      ggtgacgga ccggggcga cctccatgg gttgctgcc 
      cgagctcaa tggggtcga gctggctat gagcttgag 
      tcccaagca tcatcatca aggtggacc gcaagtctt 
      gagccatct ccagcagtt gcatcattc gacgccgtt 
      cagtgttac cgatatttt gagtcccat cgaggagtc 
      gtacaggaa attggagtt ggcggccaa tccgccgtc 
      gccctacag gtctacatc cagcggaag tagtatcaa 
      caactcccc cggagccgg tggcactcc cggaggcgg 
      cggctgcag catgaacag cagttatcc cgtaatggg 
      gcagttcgg cggtggcca atactgccc agtgtccga 
      aaacctagg tctcacccc ccactattc atcaggatg 
      gtaccaaag ctcggcagc agacccccg atttgcaat 
      ttcccgctt aatgggcag ctctgcagc ctcggcgaa 
      catgggtca cgggctgaa caccggcat gccgggcct 
      gggcgcctg cgccatgcc cgactccaa gcccatgca 
      gttcccctt ggcccaacg gcggaagag gcgggtcct 
      cttcacgca agcacaggt ttacgagct cgagaggag 
      gttcaagca gcagaagta tctgtcagc ccccgaaag 
      ggagcatct ggcctcgtt gattcatct cacgcctac 
      ccaggtgaa aatctggtt ccagaacca ccggtacaa 
      gtgcaaaag acaagccaa agaaaaggc gatggccga 
      acaaaacca acataatca gtcctccag ctcgccgcg 
      tcgggtggc cgtgcccgt gctggtcaa agacgggaa 
      gccctgctc cgccggctc ggggagcac ccagacctc 
      ctcttcagc gacctcgtc cgacacatc tcgcaccca 
      acaatcccc cgtctgcca gccccaaca acaggccca 
      atgcatggg caacggcag cttggccat ggcataccg 
      acaacaaaa caactacca acaacaatg tgggactta 
      tctgcccct tcaaacaag ggcgtggta gtgacaccg 
      tgaataaaa aattgagcg atacttaat catacatag 
      tcgaatttt tgggtcata tctcgtcct gttccttac 
      taatccggt gattgttga gagtgtgtg tggtgtatt 
      atatacagg gtgttcctc tttccccca aaatttgca 
      ataatccgt gcaattgcg gaccggaca gcgcacggt 
      tactgcagt gtagactgt acattaggt gtacaattt 
      attgttatt aattgtaca aacaaaata cacgtttta 
      tgtaaatat taggtaata tttataaat gtagtgatg 
      taaaaaagt cactcgtat gtacagatt cttagggtt 
      ataatggaa aatgcaccg tcatgtcat gtaactgtg 
      tttttaaat tcctcctca ataaaaatg tataaagca 
      t
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY