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Gene information
General information
TC032309
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.

TC003318
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_007418.3: 1199667..1201612
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
    TC032309
    iBeetle screen iB_06885

    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: 30.0% (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


    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 not developed/not fertilized - Penetrance (percentage of animals/eggs on slide): more than 80%

    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): 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
    embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): more than 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
    • >TC032309 RA
      MISPGSSPR CVRRNIAPG TLPPPLPPR KNSPTHEQT 
      ILPTTSNLS LRNGLHSTN GHKFTNNGM NEAQSVTNL 
      THNTIEIPQ IAKSNSMLE MHRNPSSEN DNIVELSSP 
      ETIYGVVIS NCDNIIDST KSQTRHLSC PTSRNKPMS 
      NQPVKSSTT PNFSHSSSD QTGHPVYEN VNLELPPKH 
      RDRVNPLER DSTVSYENL NMDYIKKLV SEGYSKDAV 
      IKALGITRN NVDMACDIL HEFGTKLG
    CDS sequence
    • >TC032309 RA
      atgatatcg ccggggagc agcccccgc tgcgtgcgt 
      cggaacatc gcccccggg acactgccg cccccgctg 
      cccccacgg aaaaactcg cccactcac gagcaaacg 
      attctgcca acgacttcg aatctctca ctacgaaac 
      ggtttacac agcacaaac gggcacaaa ttcactaac 
      aacggcatg aacgaagca caatcagta acaaatcta 
      acccacaac acgatcgag atacctcag atagcgaag 
      agcaattcc atgttggag atgcacaga aacccaagt 
      tccgaaaac gataatatt gtagaatta agctcaccc 
      gagacgatt tacggtgta gttatatca aattgtgat 
      aatattata gatagcact aagagtcaa acgcgacac 
      ttgagttgt cccacatcg agaaacaag ccgatgagc 
      aatcagccg gtgaaatcg agcacaacg ccgaatttt 
      agtcactcg agcagcgac caaacgggg catcctgtc 
      tacgaaaac gtcaacttg gagctgccc ccaaagcac 
      agagacagg gtgaacccg cttgagaga gacagcacc 
      gtgtcgtac gaaaatctg aacatggac tacattaag 
      aagttggtt agtgagggt tattccaaa gatgccgtt 
      ataaaagcg ctggggatt acgcggaac aatgtcgat 
      atggcttgt gatattttg cacgaattc gggacgaag 
      ttgggatag
    mRNA sequence
    • >TC032309 RA
      taccaggag acaaaacga ggtacaaaa tacctagat 
      ttttctgta caaaattta atttaattt aaatatagg 
      tcactaaga aaactgaag aaaagcaga agcaatttc 
      ttgaaaacg atcacatta tcatatttc ataatgtac 
      ttattttct tttgtcgta aagtaaaaa gtgcttttc 
      attcacctg gtatgtgta atggcgtcg aaatgaaat 
      aaattgttc tcatgtaac gaggttttc taatgttac 
      atttttagg atgttggcg agttgtgga cttgcgctg 
      gaagtagtt tacacgctc tggcactgt cacctcgta 
      gcgacattt cagcgtcag ccagcccga tgatatcgc 
      cggggagca gcccccgct gcgtgcgtc ggaacatcg 
      cccccggga cactgccgc ccccgctgc ccccacgga 
      aaaactcgc ccactcacg agcaaacga ttctgccaa 
      cgacttcga atctctcac tacgaaacg gtttacaca 
      gcacaaacg ggcacaaat tcactaaca acggcatga 
      acgaagcac aatcagtaa caaatctaa cccacaaca 
      cgatcgaga tacctcaga tagcgaaga gcaattcca 
      tgttggaga tgcacagaa acccaagtt ccgaaaacg 
      ataatattg tagaattaa gctcacccg agacgattt 
      acggtgtag ttatatcaa attgtgata atattatag 
      atagcacta agagtcaaa cgcgacact tgagttgtc 
      ccacatcga gaaacaagc cgatgagca atcagccgg 
      tgaaatcga gcacaacgc cgaatttta gtcactcga 
      gcagcgacc aaacggggc atcctgtct acgaaaacg 
      tcaacttgg agctgcccc caaagcaca gagacaggg 
      tgaacccgc ttgagagag acagcaccg tgtcgtacg 
      aaaatctga acatggact acattaaga agttggtta 
      gtgagggtt attccaaag atgccgtta taaaagcgc 
      tggggatta cgcggaaca atgtcgata tggcttgtg 
      atattttgc acgaattcg ggacgaagt tgggatagc 
      agcatcatt agtgccgat cagtgttgc agttattag 
      caaatttcg tttaaaata attgttccc gcgacttga 
      atttgctaa taatgtgat tatgtgtcc agtcctaat 
      tctaaatcg gtgttaggc aaatcagtt ttctgatcg 
      tcaatgtta ctgtgtttt gacggtaaa tagttcaca 
      cagtacctt aaatatgtt tttttcggt cgaaaaatc 
      gtgatctgg aactgttat ttttatagt ccttttagc 
      aaacttgga gagtatttt tgttaagga ccttgggtg 
      ttgttactc ctatggatt aatgtcata ccagttgac 
      tagccaata cgatgcaat attttgcta tgctttaaa 
      aagtatttt aacatatct atgtattta gtcatttta 
      tgaaaaaca tacgcttcg attttattt ttactaaca 
      atttaaggc gcgaacata agtctgatc actttattt 
      gatactgaa tagtacaac ttacgacgt ataattatt 
      acaaatatt ttaccttat tcgtaaact tctacagta 
      ataattact gattaattc cgataccgg tgttttttt 
      actatatga aacgtgtaa atatattgt ttcctcagg 
      aaagcgtca aaccaaagt ctgtatata gatgattga 
      aatgaatta ttatatgtt ttttattca tgtaaatat 
      ttcgccatg taattctta tgtataatt aaataaatt 
      ataataacc att
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY