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Gene information
General information
TC016225 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_007425.3: 4483071..4486863
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
    TC016225
    iBeetle screen iB_05978

    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


    Fertility
    eggs lost during procedure: potentially due to defects in egg-shells egg-morphology or egg-size, (14 dpi)
    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: < 20 embryo/egg not developed/not fertilized - Penetrance (percentage of animals/eggs on slide): more than 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    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
    • >TC016225 RA
      MATVKQVGD YKLGALIIE GKTKQVYEL PGSPDLCLL 
      LNKDRITAG DGVKAHDLA GKAAISNKT NGKVFEILN 
      SVGVKTSFV KAASDVAFI SKKCEMIPI EWVTRRLAT 
      GSFLKRNPG VPEGYRFFP PKHETFFKD DANHDPQWS 
      SEQIISAKF KVNGLEIGP TEVDIMTRM SILVFEILE 
      KVWASRNCA LIDMKIEFG VDKSGEILV ADVIDSDSW 
      RLWPSGDKR LMVDKQVYR NLTTVTSSD LDTVKRNFE 
      WIVDQLDHL APPNDHLVV ILMGSPSDA DHCKTIQKH 
      CEKLGLNTE LRVTSAHKG TEETIKIIR YYESLPLKV 
      VFIAVAGRS NGLGPVVSG NSPFPVINC PPLKAENVD 
      RDVWSSLNV PSGLGCSTV LYPEAAALS AANIIGLGN 
      YVVWSRLRV RQLDNYVTL AKADKGVKN N
    CDS sequence
    • >TC016225 RA
      atggcgacg gtcaaacag gtcggtgac tacaaattg 
      ggggccctc atcatcgag ggcaagacc aagcaagtg 
      tacgagttg ccgggatcc cccgatttg tgccttttg 
      ctgaataag gaccggatc acggcaggc gatggcgtc 
      aaagcgcac gatctggcg ggaaaggcg gcaatttcg 
      aataagaca aatggaaaa gttttcgaa attttaaat 
      tcagtcggg gtgaaaact tcgttcgtt aaagccgcc 
      tctgatgtg gcttttatt tcgaaaaag tgtgagatg 
      atccccatt gagtgggtg acaaggcgt ctggcgact 
      ggttccttc ctgaagcgg aatccgggg gttcccgag 
      ggatacagg tttttcccc ccgaaacac gagactttt 
      ttcaaagat gacgcaaac cacgacccc caatggtcc 
      agcgagcag attatctcg gcgaagttc aaggtcaat 
      ggcttggag attgggccc actgaggtt gatatcatg 
      acgcggatg tcgattttg gtttttgaa attttggaa 
      aaagtctgg gcgtctaga aattgcgct cttatcgat 
      atgaaaata gaattcggt gtcgacaaa tcaggggaa 
      atcctcgtg gccgacgtc atcgactcg gacagctgg 
      cggctgtgg ccctccggg gacaagcgc ctcatggtc 
      gacaagcaa gtctaccgt aacctcacc accgtcacc 
      tcctccgac ctggacacc gtcaagcgc aacttcgag 
      tggatcgtc gaccagctg gaccacctc gccccgccc 
      aacgaccac ctggtggtc atcctcatg ggcagcccc 
      tcggacgcc gaccactgc aaaaccatc cagaaacac 
      tgcgaaaaa ctcggtctc aacaccgaa ctgcgcgtc 
      acctcggca cacaaaggc acagaagaa acgatcaaa 
      attatcaga tattacgag tctctccct ctgaaggtc 
      gtttttatc gcagtggcg ggccggtcc aacggcttg 
      ggaccagtg gtttcgggg aattccccg ttcccggtc 
      attaactgt cccccactt aaggccgaa aatgtggac 
      agggacgtg tggtccagc ttgaatgtg ccctcgggg 
      ctggggtgc agtacggtc ctgtatccc gaagctgcc 
      gccttaagt gcggccaat attatcggg ctgggaaat 
      tatgtggtc tggagtagg ctgagagtg agacagttg 
      gataattat gtcactctg gccaaggct gataagggg 
      gtgaagaat aattga
    mRNA sequence
    • >TC016225 RA
      gcacgaatc atctacgat ttataaaaa caaacaacg 
      gtcgctttt cgtcctctt gcacgtttt ctgcaagaa 
      ttgtgtttt tgttgccat ggcgacggt caaacaggt 
      cggtgacta caaattggg ggccctcat catcgaggg 
      caagaccaa gcaagtgta cgagttgcc gggatcccc 
      cgatttgtg ccttttgct gaataagga ccggatcac 
      ggcaggcga tggcgtcaa agcgcacga tctggcggg 
      aaaggcggc aatttcgaa taagacaaa tggaaaagt 
      tttcgaaat tttaaattc agtcggggt gaaaacttc 
      gttcgttaa agccgcctc tgatgtggc ttttatttc 
      gaaaaagtg tgagatgat ccccattga gtgggtgac 
      aaggcgtct ggcgactgg ttccttcct gaagcggaa 
      tccgggggt tcccgaggg atacaggtt tttcccccc 
      gaaacacga gactttttt caaagatga cgcaaacca 
      cgaccccca atggtccag cgagcagat tatctcggc 
      gaagttcaa ggtcaatgg cttggagat tgggcccac 
      tgaggttga tatcatgac gcggatgtc gattttggt 
      ttttgaaat tttggaaaa agtctgggc gtctagaaa 
      ttgcgctct tatcgatat gaaaataga attcggtgt 
      cgacaaatc aggggaaat cctcgtggc cgacgtcat 
      cgactcgga cagctggcg gctgtggcc ctccgggga 
      caagcgcct catggtcga caagcaagt ctaccgtaa 
      cctcaccac cgtcacctc ctccgacct ggacaccgt 
      caagcgcaa cttcgagtg gatcgtcga ccagctgga 
      ccacctcgc cccgcccaa cgaccacct ggtggtcat 
      cctcatggg cagcccctc ggacgccga ccactgcaa 
      aaccatcca gaaacactg cgaaaaact cggtctcaa 
      caccgaact gcgcgtcac ctcggcaca caaaggcac 
      agaagaaac gatcaaaat tatcagata ttacgagtc 
      tctccctct gaaggtcgt ttttatcgc agtggcggg 
      ccggtccaa cggcttggg accagtggt ttcggggaa 
      ttccccgtt cccggtcat taactgtcc cccacttaa 
      ggccgaaaa tgtggacag ggacgtgtg gtccagctt 
      gaatgtgcc ctcggggct ggggtgcag tacggtcct 
      gtatcccga agctgccgc cttaagtgc ggccaatat 
      tatcgggct gggaaatta tgtggtctg gagtaggct 
      gagagtgag acagttgga taattatgt cactctggc 
      caaggctga taagggggt gaagaataa ttgaacaaa 
      tgccatttt ttatttcgt gttttattt aaaaaaata 
      cataaaaag taataat
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY