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Gene information
General information
TC003323 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: 1238517..1240279
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
    TC003323
    iBeetle screen iB_09732

    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: 70.0% (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: 4 (9 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 2 (11 dpi)
    Days post injection


    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
    • >TC003323 RA
      MAANITKVP LLRAVATRG YASCPPAPI GGIHDYEVK 
      NTTLPNNLV VASAENECP ISRISIVFR AGSRNETHE 
      NAGVTHTLR ICAGLSTKN ATQFAITRN IQQAGATLT 
      ATSDREIVS YTLEGTRKA VEKTLPFLT EVATQQVFK 
      PWEVSENVG RQRLELAIR PPQLRAIDL VHKAAFRRG 
      LGNSLYSAK YNLGNISSE TLQHYVASN FLSGRAAVV 
      GLGVDHSQL VKYAQGLAL ESGEGTSNP SPYFGGEIR 
      SDKGGDFAY VAIAGQGAP WKNSKEALA VSVLQKALG 
      GGPKVKWGS VDNGALSKV VGGEGDAKY ALNTFNASY 
      SDAGIFGVL IAAPEATAG KIVQAAFKL LKAGNLTDA 
      DVNRGKNQL KAALLIKNE SGSSAIDFL GSQAAVLGS 
      AKSPSQVVA EIDSITTAD VNAALKKVA SGKLSIASV 
      GQLRTVPFL DELK
    CDS sequence
    • >TC003323 RA
      atggctgca aatatcaca aaggttccc ctgcttcga 
      gccgtagcg acacgtggc tacgccagc tgcccccca 
      gcccccatc ggaggcatc cacgactac gaagtcaaa 
      aataccact ctacccaac aatttggtc gtggcctcg 
      gccgaaaac gaatgcccc atttcgcgc atttccatc 
      gtctttagg gccggttcc cgcaacgaa acgcatgaa 
      aatgccggc gtgacacac actcttcgc atttgcgct 
      ggcttatca acgaaaaat gccacacag tttgccatt 
      acacgtaac atccagcaa gctggggcc accttgacg 
      gccacctcc gatcgcgag atcgtttcg tacactttg 
      gagggcacc aggaaagcc gtcgagaaa acgcttccg 
      ttcctcaca gaagtggcc actcagcag gttttcaaa 
      ccatgggaa gtgtcagaa aatgtcggt cgccaaagg 
      cttgaactc gccattcgg ccaccacaa ttgagggca 
      attgatttg gttcacaaa gctgcgttc cgtcgcggt 
      cttggaaac tccctctac agtgctaaa tacaacttg 
      gggaacatt tcatctgag actctccaa cattacgtt 
      gccagtaat ttcctttcc ggacgtgct gctgttgtt 
      ggtttggga gtagatcac tcacaactt gttaaatac 
      gcccaagga ctagccctt gaaagtggg gaaggtaca 
      tccaatccc tctccgtac tttgggggc gaaatcagg 
      tctgacaaa gggggcgat tttgcatac gttgcgatc 
      gccggccaa ggcgcccca tggaagaat tccaaagaa 
      gcgcttgct gtttcagtc cttcagaaa gccctaggc 
      ggaggcccc aaagtcaag tgggggtct gtagacaat 
      ggggccttg agtaaagtt gttggaggc gaaggcgat 
      gccaaatac gccctcaat acgttcaat gccagctac 
      agtgatgcc gggattttt ggtgtcttg attgcagca 
      ccggaagcc acagccggg aaaattgtg caagcagca 
      ttcaaattg ctaaaagct gggaatttg accgatgcg 
      gatgttaac agagggaag aatcaattg aaagcggcg 
      cttcttatt aaaaatgag tctggttca agcgctatt 
      gattttctc ggaagtcag gcggctgtt ttggggtcg 
      gcgaaatcg ccaagtcag gtggtggca gagattgat 
      tcaatcaca acagcagat gtcaatgct gctctcaag 
      aaagttgct tcaggtaaa ctatcaata gcctccgtt 
      ggccaactg cgcaccgta ccattttta gatgaacta 
      aagtaa
    mRNA sequence
    • >TC003323 RA
      agggacttt tttgtgaaa ttacgtaac tgacatttg 
      acagttgtg gccgctgat gtccggaac ggaaattca 
      ataagagat attttcaaa aactcttat ttaaaccaa 
      ttgctctca aatcacaat ggctgcaaa tatcacaaa 
      ggttcccct gcttcgagc cgtagcgac acgtggcta 
      cgccagctg ccccccagc ccccatcgg aggcatcca 
      cgactacga agtcaaaaa taccactct acccaacaa 
      tttggtcgt ggcctcggc cgaaaacga atgccccat 
      ttcgcgcat ttccatcgt ctttagggc cggttcccg 
      caacgaaac gcatgaaaa tgccggcgt gacacacac 
      tcttcgcat ttgcgctgg cttatcaac gaaaaatgc 
      cacacagtt tgccattac acgtaacat ccagcaagc 
      tggggccac cttgacggc cacctccga tcgcgagat 
      cgtttcgta cactttgga gggcaccag gaaagccgt 
      cgagaaaac gcttccgtt cctcacaga agtggccac 
      tcagcaggt tttcaaacc atgggaagt gtcagaaaa 
      tgtcggtcg ccaaaggct tgaactcgc cattcggcc 
      accacaatt gagggcaat tgatttggt tcacaaagc 
      tgcgttccg tcgcggtct tggaaactc cctctacag 
      tgctaaata caacttggg gaacatttc atctgagac 
      tctccaaca ttacgttgc cagtaattt cctttccgg 
      acgtgctgc tgttgttgg tttgggagt agatcactc 
      acaacttgt taaatacgc ccaaggact agcccttga 
      aagtgggga aggtacatc caatccctc tccgtactt 
      tgggggcga aatcaggtc tgacaaagg gggcgattt 
      tgcatacgt tgcgatcgc cggccaagg cgccccatg 
      gaagaattc caaagaagc gcttgctgt ttcagtcct 
      tcagaaagc cctaggcgg aggccccaa agtcaagtg 
      ggggtctgt agacaatgg ggccttgag taaagttgt 
      tggaggcga aggcgatgc caaatacgc cctcaatac 
      gttcaatgc cagctacag tgatgccgg gatttttgg 
      tgtcttgat tgcagcacc ggaagccac agccgggaa 
      aattgtgca agcagcatt caaattgct aaaagctgg 
      gaatttgac cgatgcgga tgttaacag agggaagaa 
      tcaattgaa agcggcgct tcttattaa aaatgagtc 
      tggttcaag cgctattga ttttctcgg aagtcaggc 
      ggctgtttt ggggtcggc gaaatcgcc aagtcaggt 
      ggtggcaga gattgattc aatcacaac agcagatgt 
      caatgctgc tctcaagaa agttgcttc aggtaaact 
      atcaatagc ctccgttgg ccaactgcg caccgtacc 
      atttttaga tgaactaaa gtaaactta agtcggctt 
      aatttatgt tgtataaat tctgtttta aactaaact 
      tatacatgt tag
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY