Status
Please wait ...
Gene information
General information
TC032266
Former TC numbers

If one gene is splitted or genes are merged, the original TC numbers are listed. One listed former TC number indicates the previous gene has be splitted, serveral former TC number represents a merged gene.

TC000189,TC000190
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_007417.3: 14937469..14939249
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
    TC032266
    iBeetle screen iB_09709

    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: 20.0% (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


    adult lethality - Number of animals showing the phenotype: 1 (9 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 embryo/egg with embryonic tissue, no cuticle, no fluorescence - 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): 30% - 50%

    embryo/egg with muscle fluorescence/pattern, but not hatched (pot. late embryonic lethality) - 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 - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    remarks: muscle pattern potentially obscured by segmentation defect,
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): less than 30%
    cuticle remnants with unidentifyable segments - Penetrance (percentage of animals/eggs on slide): less than 30%

    larva constricted - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    Comment: Constriction often occurs between thorax and abdomen segments.
    head appendages position irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    head not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    head shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    head not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    thorax not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    thorax shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    thorax not present - Penetrance (percentage of animals/eggs on slide): less than 30%
    abdomen not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    abdomen shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    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
    • >TC032266 RA
      MVIISKPLF YDKLELAIS CRELVHPDV GSCTKANLK 
      LCSEDFLKG VFYFYLPIH LLQVILDYR KIKRKHFLK 
      NLLQNVLRS ECFGVSFGA LIIATICLQ RHFTERFHY 
      YSLLLLPGV VAGISLRWE HPRNRQLNT ILWASLVVE 
      ALFKNAYYF KIFSYSRIK ETLIFMIFS SLLMHLFVN 
      RDKKHIPFN LWFYKPSHP KKFKFTLWN YFIEISKYF 
      GFGYGLNSV RCVLKDAKM VFKNPLQFV RKTIFNRRN 
      VKFGIIIGA YVGLFRGVS VLLQKYTDF ERANAAIAG 
      LVAGLSYYI QPNMTVLTT AIVTVFQIL CQQIATKPK 
      FRKIPFAEL IFMFCNGIL FFNRIVAPV TCPKYYMNM 
      LRITGNGEL EKIYVRLVN KYLP
    CDS sequence
    • >TC032266 RA
      atggttatt ataagcaaa ccgctattt tacgataaa 
      cttgagctt gcaatatcg tgccgggag ttagttcat 
      ccagatgtt ggcagttgt acgaaagcg aatctcaaa 
      ctttgttcg gaggatttt ttaaaagga gtgttttat 
      ttctattta cccattcat ttgttgcaa gttattctc 
      gattataga aaaatcaag cgcaagcat tttctcaaa 
      aatctgcta cagaacgtc ctaaggtct gaatgtttc 
      ggtgtcagt tttggggca ctgataatt gccacaatc 
      tgtctacaa cgtcatttt acagaacgg ttccattat 
      tattctcta ttattactg ccaggtgta gtggcagga 
      atttctcta cgatgggaa caccctaga aatcggcag 
      ttaaatacg attctttgg gccagtttg gttgttgag 
      gcattgttc aaaaatgct tactatttc aaaatattc 
      agttattct cggataaaa gaaacgctt atttttatg 
      atttttagt tctcttttg atgcattta tttgtaaac 
      agggataag aaacacatt ccgttcaat ctttggttt 
      tataaacca tcccacccg aaaaaattc aaattcacg 
      ttgtggaat tattttatc gaaatttca aagtatttc 
      ggattcggc tatggatta aatagtgtt aggtgcgtc 
      ttgaaagac gccaaaatg gtgtttaaa aaccctctg 
      caatttgtt agaaaaacg atctttaac agaagaaat 
      gttaaattt ggaataatt attggtgct tacgttggt 
      ttatttagg ggggtttca gttttactt cagaaatat 
      acggacttt gaaagagca aatgctgca attgcagga 
      cttgtagca ggattatct tactatatt cagccaaat 
      atgaccgtt ttgaccacc gctattgta actgttttt 
      caaatatta tgtcagcaa attgcaacc aaaccaaag 
      tttcgaaaa attccgttt gctgaatta atttttatg 
      ttttgtaat gggattctt ttcttcaat agaattgta 
      gctccagtt acttgcccg aaatattac atgaatatg 
      cttcgaatt actggaaat ggagaactg gaaaaaatc 
      tacgttaga ctagtgaac aagtatttg ccataa
    mRNA sequence
    • >TC032266 RA
      taagtattt attcggtgt gttggaaac aatagtctg 
      aataataaa aactaccaa cgacattat ttcgctgct 
      tgagtccag taagcaaac atgaaagaa aaaagcgct 
      tgaactgta ataattgtg actggtggc aaaatggtt 
      attataagc aaaccgcta ttttacgat aaacttgag 
      cttgcaata tcgtgccgg gagttagtt catccagat 
      gttggcagt tgtacgaaa gcgaatctc aaactttgt 
      tcggaggat tttttaaaa ggagtgttt tatttctat 
      ttacccatt catttgttg caagttatt ctcgattat 
      agaaaaatc aagcgcaag cattttctc aaaaatctg 
      ctacagaac gtcctaagg tctgaatgt ttcggtgtc 
      agttttggg gcactgata attgccaca atctgtcta 
      caacgtcat tttacagaa cggttccat tattattct 
      ctattatta ctgccaggt gtagtggca ggaatttct 
      ctacgatgg gaacaccct agaaatcgg cagttaaat 
      acgattctt tgggccagt ttggttgtt gaggcattg 
      ttcaaaaat gcttactat ttcaaaata ttcagttat 
      tctcggata aaagaaacg cttattttt atgattttt 
      agttctctt ttgatgcat ttatttgta aacagggat 
      aagaaacac attccgttc aatctttgg ttttataaa 
      ccatcccac ccgaaaaaa ttcaaattc acgttgtgg 
      aattatttt atcgaaatt tcaaagtat ttcggattc 
      ggctatgga ttaaatagt gttaggtgc gtcttgaaa 
      gacgccaaa atggtgttt aaaaaccct ctgcaattt 
      gttagaaaa acgatcttt aacagaaga aatgttaaa 
      tttggaata attattggt gcttacgtt ggtttattt 
      aggggggtt tcagtttta cttcagaaa tatacggac 
      tttgaaaga gcaaatgct gcaattgca ggacttgta 
      gcaggatta tcttactat attcagcca aatatgacc 
      gttttgacc accgctatt gtaactgtt tttcaaata 
      ttatgtcag caaattgca accaaacca aagtttcga 
      aaaattccg tttgctgaa ttaattttt atgttttgt 
      aatgggatt cttttcttc aatagaatt gtagctcca 
      gttacttgc ccgaaatat tacatgaat atgcttcga 
      attactgga aatggagaa ctggaaaaa atctacgtt 
      agactagtg aacaagtat ttgccataa aatcaaata 
      gctaattag ttaattttt gtttacttt tttaacact 
      aataaacct ctaacataa at
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY