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Gene information
General information
TC008454 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_007419.2: 10755715..10761139
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
    TC008454
    iBeetle screen iB_01351

    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: 40.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 4 (3 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature fluorescence degenerated - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): less than 30%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    head slightly posterior segment boundary irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 30% - 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: 30.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 30.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality early (L5/6) - Number of animals showing the phenotype: 2 (11 dpi)
    Days post injection


    Fertility
    egg production: number <50% of expected -
    mature egg deposition blocked number 1st lateral oviduct: 2 number 2nd lateral oviduct: - Number of animals showing the phenotype: 2
    ovary thinner - Number of animals showing the phenotype: 2
    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC008454 RA
      MEVVMHEAA NPTEESESV KYNSAEKSV TLMQEEESG 
      TFSRPPQQT IDLQFEDIK YTASLGFRK GTKEILHGV 
      NGKFPPGQL IAIMGPSGA GKSTLLDIL SGYRIRGVS 
      GTVYANGFP RNLKAFRKS SCYIQQDDR LQPLLTTAE 
      NMQIAADLK LGPEVSQRE KTETIEEIL KWLGLSKTM 
      NTKAAGLSG GQKKRLSIA LELINNPTV MFLDEPTTG 
      LDSSSCSQC IKLLKDLAR QGRTIVCTI HQPSASLFQ 
      LFDQVYVLA AGRCLYQGS TTNLVPFLN DVKYPCPQY 
      HNPADFVIE LACEEYGPE IIDTMVATT ENGSSVAYF 
      DKEVVPTLK RPIAGGNSA KTSSDDESG LQATSSYNQ 
      LKVLLRRGY IKTKRDQTL TYMRLMVNA SVGLMLGTL 
      YWQAGSDGT KVLDNYNLL FSILMHHMM STMMLTILT 
      FPNEMSILI KEHFNRWYS LKMYYTSVT LVDIPVSIA 
      CCALFSTII YYMTGQPLD KQRFMMFFV ISMLVVFVA 
      QSFGLMVGA VFNVINGTF LGPTLSVPM MMFAGFGVT 
      LRDLPGYLY WGSYISYLR YGLEGIVGA IYGLERETI 
      WCPDDAYCH YKYPKKLLE DIAMKGDQF GNDVIALVL 
      ILFFLRIVA YVLLRYKLM AVR
    CDS sequence
    • >TC008454 RA
      atggaagtt gtaatgcac gaagcggcc aacccgacg 
      gaggaaagt gagagtgtt aaatataac agtgcggaa 
      aaatcagtg actttgatg caagaggaa gaaagtggt 
      actttttcc agaccgccc cagcaaacc atcgactta 
      cagttcgaa gatatcaaa tacacggcc tctttgggg 
      tttagaaaa ggtactaag gaaatcctc catggagtc 
      aatgggaaa tttccccca ggccagctc atcgcaatt 
      atgggcccg tcaggggct ggtaaaagt accctcctt 
      gacatcctc agcgggtac cgcatccgt ggcgtatct 
      ggcacagtt tatgccaac ggttttccc cgaaacttg 
      aaagcgttt cgcaagtcg tcttgctac atccagcaa 
      gacgaccgg ctccaaccc ctcttgacc acagcggag 
      aacatgcaa atcgccgcc gatctcaaa ctaggccct 
      gaagtttcg caacgggag aagaccgaa acgattgaa 
      gaaattttg aaatggttg ggattgagc aagacgatg 
      aacacgaag gcggcgggg ttgtccgga ggccagaag 
      aagcggctg tccatcgct ttggagctt ataaataac 
      ccaactgtt atgtttctt gatgagcca acaaccggt 
      ttggacagc tcttcctgt tcccaatgc atcaaactg 
      ctcaaggat ttggcgcgc caagggagg accatcgtg 
      tgtaccatc catcaacca tcggcctcc cttttccag 
      ctattcgac caagtctac gtactggcc gctgggcgg 
      tgtttgtac caaggcagt accacaaat ctagtcccg 
      tttctgaac gacgtcaag tacccatgt ccacaatac 
      cataaccca gccgatttt gtcatcgaa ttggcgtgc 
      gaagagtac gggcctgaa ataatcgac accatggtc 
      gcaaccacc gagaacggc agttcggtg gcgtacttc 
      gacaaagaa gtagtacca acattgaaa cgaccaatc 
      gctggaggc aattcggcg aaaacatcc tccgatgat 
      gaatccggg ttacaagcg accagttcg tacaatcag 
      ttgaaagtg ttgttacga aggggatac attaagaca 
      aaacgtgat cagactttg acgtatatg cggctcatg 
      gtcaatgct tctgttggt ctaatgttg ggaacgttg 
      tactggcag gcggggtca gatggtacc aaagtttta 
      gataattat aatttgctc ttctccata cttatgcat 
      cacatgatg tccacgatg atgctcacc atattgaca 
      tttcccaac gaaatgtcc attttgata aaagagcac 
      ttcaaccgg tggtactcc ctcaaaatg tattacact 
      tcagtaacc ctagtcgac attccagtt tcgatcgca 
      tgttgtgcc ctcttttcg acaataatt tactacatg 
      accggacaa ccccttgac aaacaacgc tttatgatg 
      tttttcgtc ataagtatg ctcgtggtt ttcgtggcc 
      caaagtttc ggtttaatg gttggtgcc gtcttcaat 
      gttattaac ggcacattt ttggggccc actctatcc 
      gttcctatg atgatgttt gccgggttt ggggtcaca 
      cttcgagac ctccctgga tatctctac tgggggagt 
      tatatttcg tatctgaga tacggactt gaagggatt 
      gttggggcg atttacggg cttgaaagg gagacgatt 
      tggtgtcca gacgatgcc tactgtcat tataaatac 
      ccgaagaaa ctgcttgag gatattgca atgaagggc 
      gaccagttt ggaaacgat gtaatcgca ctggttctc 
      attttgttc tttttgagg atcgttgcg tacgttctt 
      ctgcggtac aagttaatg gcggttcgc tga
    mRNA sequence
    • >TC008454 RA
      tcacagacc agaaaaaac cagtggggc cttgaacta 
      ggtaccata cctgatgca taattaatt tctcaaaat 
      accagttag ttgtgacga aaccgctga attgaattt 
      atttgaaaa aattgctga ttgagtttg tgcacattc 
      gaaaagcga gcgagcatc tcgtttgtg acaatacga 
      gtactctca cacgagagt ggactttaa tcgcggtcc 
      aatctgttt aactttcca cactctata taagtgacc 
      aaagcaaga tgataataa ttcactaga aataagcgc 
      gatcgcaaa caacttttg ttcattctt ccatgtcgg 
      tgtaaagtg aacacgtct caccatgga agttgtaat 
      gcacgaagc ggccaaccc gacggagga aagtgagag 
      tgttaaata taacagtgc ggaaaaatc agtgacttt 
      gatgcaaga ggaagaaag tggtacttt ttccagacc 
      gccccagca aaccatcga cttacagtt cgaagatat 
      caaatacac ggcctcttt ggggtttag aaaaggtac 
      taaggaaat cctccatgg agtcaatgg gaaatttcc 
      cccaggcca gctcatcgc aattatggg cccgtcagg 
      ggctggtaa aagtaccct ccttgacat cctcagcgg 
      gtaccgcat ccgtggcgt atctggcac agtttatgc 
      caacggttt tccccgaaa cttgaaagc gtttcgcaa 
      gtcgtcttg ctacatcca gcaagacga ccggctcca 
      acccctctt gaccacagc ggagaacat gcaaatcgc 
      cgccgatct caaactagg ccctgaagt ttcgcaacg 
      ggagaagac cgaaacgat tgaagaaat tttgaaatg 
      gttgggatt gagcaagac gatgaacac gaaggcggc 
      ggggttgtc cggaggcca gaagaagcg gctgtccat 
      cgctttgga gcttataaa taacccaac tgttatgtt 
      tcttgatga gccaacaac cggtttgga cagctcttc 
      ctgttccca atgcatcaa actgctcaa ggatttggc 
      gcgccaagg gaggaccat cgtgtgtac catccatca 
      accatcggc ctccctttt ccagctatt cgaccaagt 
      ctacgtact ggccgctgg gcggtgttt gtaccaagg 
      cagtaccac aaatctagt cccgtttct gaacgacgt 
      caagtaccc atgtccaca ataccataa cccagccga 
      ttttgtcat cgaattggc gtgcgaaga gtacgggcc 
      tgaaataat cgacaccat ggtcgcaac caccgagaa 
      cggcagttc ggtggcgta cttcgacaa agaagtagt 
      accaacatt gaaacgacc aatcgctgg aggcaattc 
      ggcgaaaac atcctccga tgatgaatc cgggttaca 
      agcgaccag ttcgtacaa tcagttgaa agtgttgtt 
      acgaagggg atacattaa gacaaaacg tgatcagac 
      tttgacgta tatgcggct catggtcaa tgcttctgt 
      tggtctaat gttgggaac gttgtactg gcaggcggg 
      gtcagatgg taccaaagt tttagataa ttataattt 
      gctcttctc catacttat gcatcacat gatgtccac 
      gatgatgct caccatatt gacatttcc caacgaaat 
      gtccatttt gataaaaga gcacttcaa ccggtggta 
      ctccctcaa aatgtatta cacttcagt aaccctagt 
      cgacattcc agtttcgat cgcatgttg tgccctctt 
      ttcgacaat aatttacta catgaccgg acaacccct 
      tgacaaaca acgctttat gatgttttt cgtcataag 
      tatgctcgt ggttttcgt ggcccaaag tttcggttt 
      aatggttgg tgccgtctt caatgttat taacggcac 
      atttttggg gcccactct atccgttcc tatgatgat 
      gtttgccgg gtttggggt cacacttcg agacctccc 
      tggatatct ctactgggg gagttatat ttcgtatct 
      gagatacgg acttgaagg gattgttgg ggcgattta 
      cgggcttga aagggagac gatttggtg tccagacga 
      tgcctactg tcattataa atacccgaa gaaactgct 
      tgaggatat tgcaatgaa gggcgacca gtttggaaa 
      cgatgtaat cgcactggt tctcatttt gttcttttt 
      gaggatcgt tgcgtacgt tcttctgcg gtacaagtt 
      aatggcggt tcgctgatt tagctctaa gttatttat 
      ttgtgtaag gctgtgtta ttttatttt tatatttat 
      taaatgtgt gttgaatta
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