Status
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Gene information
General information
TC032664
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.

TC003974,TC003975
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: 29548724..29576877
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
    TC032664
    iBeetle screen iB_03662

    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: 3 (3 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): 30% - 50%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    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): 50% - 80%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 50% - 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: 0.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 10.0% (includes death as larva, prepupa, pupa, adult)

    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (16 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 4 (41 dpi)
    Days post injection


    Adult morphology
    elytra dorsal orientation not closed - Number of animals showing the phenotype: 3


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC032664 RA
      MLKERESIE RAISSPCSS LGGSSTLDG TTCAGCGGR 
      IQDRYYLLA VDRQWHASC LKCCECKLP LDTELTCFA 
      RDGNIYCKE DYYRMFAVT RCGRCQAGI SANELVMRA 
      RDSVYHLHC FSCTSCGIP LSKGDHFGM RDGLIYCRP 
      HYELLDFCD TSDPVEMMF RGSESPPGY FANAAPQHH 
      KGRPRKRKL PVEDRGCGE IPVTMRMAA ATLEMLQQG 
      DLSSSMESL SYDSSVTSP ASSNGHQPQ RTKRMRTSF 
      KHHQLRTMK TYFAINQNP DAKDLKQLA QKTGLSKRV 
      LQVWFQNAR AKWRRNIMR QEGGQSQGQ SQANGPPSV 
      GSGIMGEGN SLSHQSLDD LHHHLHSQN SQTLTFSDI 
      Y
    CDS sequence
    • >TC032664 RA
      atgctgaag gagcgcgaa tccatcgag cgtgccatc 
      tcctcgccc tgctccagc cttggcggc tcctccacg 
      ctggacggg acgacctgt gcgggctgc ggaggccga 
      atccaggac cgttactac ctcctggcc gtcgacaga 
      caatggcat gcctcctgc ttgaaatgt tgcgaatgt 
      aaacttcct ctggatacg gagctgact tgtttcgcc 
      cgagatggg aacatttac tgcaaggaa gattattat 
      cggatgttt gcggtgacg cgctgtggc cgctgccag 
      gcgggcatc tccgccaac gagctggtg atgcgagcc 
      agagactcc gtctaccac ctgcactgc ttctcgtgc 
      acctcctgc gggatcccg ctgtccaaa ggagaccac 
      ttcgggatg agagatgga ctcatttat tgtcggcca 
      cattacgag ctgctggat ttttgtgat acgagcgat 
      ccggtggag atgatgttc agagggagc gagtcgcca 
      ccggggtat tttgccaac gcggcgcca cagcaccac 
      aaggggaga cccaggaag cggaaactg ccggtggag 
      gatcggggg tgtggggaa attccggtc acaatgaga 
      atggcagcg gcaacacta gagatgctc cagcaaggc 
      gacctctcc tcgtccatg gagtcgctc tcgtacgac 
      tcgtcggtg acgtccccc gcgtccagc aacggccac 
      cagccccag cggaccaag cgcatgcgc acgtccttc 
      aagcaccac cagctgagg accatgaag acgtacttc 
      gccatcaat cagaacccc gacgccaag gacctgaag 
      cagctggcg cagaagacg ggcctctcg aagagggtg 
      ctccaggtg tggttccag aacgccagg gccaagtgg 
      aggaggaac atcatgcgg caggaaggc ggccaaagc 
      cagggccag agccaggcc aatgggccc ccgtccgtg 
      ggctcgggg atcatgggg gaggggaac agtttgtcg 
      catcagtcg ctagatgat ctccaccat catctgcat 
      tcgcaaaat tcgcaaact ttgacgttc agtgatatt 
      tattag
    mRNA sequence
    • >TC032664 RA
      cgacggtaa actgcacgg tggctgggc tcgaaacgt 
      cgcgagtgt tgctgtgtt gtgctgttt tgacagttc 
      ttgtggatt atgggcggc gttgccagg tagctgcag 
      cttcagtcg tttttgggc tcgtgcaag atgctgaag 
      gagcgcgaa tccatcgag cgtgccatc tcctcgccc 
      tgctccagc cttggcggc tcctccacg ctggacggg 
      acgacctgt gcgggctgc ggaggccga atccaggac 
      cgttactac ctcctggcc gtcgacaga caatggcat 
      gcctcctgc ttgaaatgt tgcgaatgt aaacttcct 
      ctggatacg gagctgact tgtttcgcc cgagatggg 
      aacatttac tgcaaggaa gattattat cggatgttt 
      gcggtgacg cgctgtggc cgctgccag gcgggcatc 
      tccgccaac gagctggtg atgcgagcc agagactcc 
      gtctaccac ctgcactgc ttctcgtgc acctcctgc 
      gggatcccg ctgtccaaa ggagaccac ttcgggatg 
      agagatgga ctcatttat tgtcggcca cattacgag 
      ctgctggat ttttgtgat acgagcgat ccggtggag 
      atgatgttc agagggagc gagtcgcca ccggggtat 
      tttgccaac gcggcgcca cagcaccac aaggggaga 
      cccaggaag cggaaactg ccggtggag gatcggggg 
      tgtggggaa attccggtc acaatgaga atggcagcg 
      gcaacacta gagatgctc cagcaaggc gacctctcc 
      tcgtccatg gagtcgctc tcgtacgac tcgtcggtg 
      acgtccccc gcgtccagc aacggccac cagccccag 
      cggaccaag cgcatgcgc acgtccttc aagcaccac 
      cagctgagg accatgaag acgtacttc gccatcaat 
      cagaacccc gacgccaag gacctgaag cagctggcg 
      cagaagacg ggcctctcg aagagggtg ctccaggtg 
      tggttccag aacgccagg gccaagtgg aggaggaac 
      atcatgcgg caggaaggc ggccaaagc cagggccag 
      agccaggcc aatgggccc ccgtccgtg ggctcgggg 
      atcatgggg gaggggaac agtttgtcg catcagtcg 
      ctagatgat ctccaccat catctgcat tcgcaaaat 
      tcgcaaact ttgacgttc agtgatatt tattagggt 
      agtgacact gtatagact agttaggtt cacacacgc 
      aatgttcct tcatgatag tgataattg gatgatttt 
      atcggttgt ttttcagtt gaggatttt gttagagga 
      tagattgca cgatgagtt ttgaggatc ttgtagata 
      atgtcgttg tttgtacat aagaatata cgttagtgg 
      taggcgaga gttttgagg aatatttat aattttgta 
      tgggtgtgc gatgatgtc atacatgtg cggtgtatg 
      tctacaagg aataaatgt atataattt at
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY