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Gene information
General information
TC004521 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: 3299181..3301137
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
    TC004521
    iBeetle screen iB_08971

    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: 60.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 4 (3 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 2 (9 dpi)
    Days post injection


    adult nutritional state potentially bad (cachectic) - Number of animals showing the phenotype: 4 (11 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: < 20 embryo/egg with embryonic tissue potentially present, no cuticle, no fluorescence - 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%
    cuticle remnants strongly fragmented (cuticle crumbs) - Penetrance (percentage of animals/eggs on slide): less than 30%

    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
    • >TC004521 RA
      MEEKPFAVE VARQGRALC KKCRQKCVQ GDLRIAKLI 
      PNPLGSGKM KTWYHVSCI FETFLKQRQ TTKRIEGPL 
      DIDGWESLD PDDKVNILE KISQCNREF EAKFSKKRP 
      KKGTKPQPQ LVVDDNLSN LSDNCTTEE VHKDNCFRE 
      FRRIVSKLP KVSSSNEKI SIFREIFTK GCDGSGFKG 
      DVVLWCKLL LPAVNKRVF HLQTKQLIK IFSKIFAVS 
      EETMTAHLE QGDVGQTIQ HFFEENAEC KPAKKSTLS 
      LFEVDDFLE SLTTLSKEE DQCNRFKTF IKKCTANDL 
      KVVIRLIKH DLRMSAGAK HVFNGLHPE AYSVFQTCK 
      DIATVISTC QGRKEPPKK RRKISEEKR KSIPNYFEN 
      VRLFLEEGV KERHAQWVK YFIAYGGSV LTNDKPEEA 
      THVLHLKDT ISQPTIHCA NSAKHVIVE WIKDTVAEE 
      NLQDYRRYT VRWDPCHKD
    CDS sequence
    • >TC004521 RA
      atggaggag aagcccttc gccgtggag gtggccagg 
      caagggcgc gccctttgc aaaaaatgc cgccagaag 
      tgcgttcaa ggcgatttg cgcattgct aaattaatc 
      ccgaaccct ctgggttcc ggtaaaatg aaaacttgg 
      taccacgta agctgtatc tttgaaaca tttttaaaa 
      cagagacaa acaactaaa agaatcgag ggccccctg 
      gacatcgac ggctgggag tcactcgac cccgacgat 
      aaagtgaat atcttggaa aagatctcg caatgtaac 
      cgggaattt gaagccaag tttagtaaa aaaagaccg 
      aaaaagggg accaagccg cagccgcag cttgtcgtc 
      gacgacaat ttgagtaat ttatctgat aattgcacg 
      actgaggag gttcacaag gataattgt ttccgcgag 
      tttcgcagg atcgtgtcg aagctcccg aaggtgtca 
      agttccaac gaaaaaatc agcattttc cgcgagatt 
      ttcacgaaa ggttgcgac gggagcggt tttaagggc 
      gatgtcgtt ctctggtgc aaactgctc ctccctgcg 
      gtgaacaaa cgcgtgttt cacttgcag accaagcaa 
      ctgattaaa atatttagc aagattttc gcggttagt 
      gaagaaaca atgactgct cacttggaa cagggcgat 
      gtgggccag accatccaa catttcttc gaagaaaac 
      gccgagtgt aaaccggcg aaaaaaagc accttgagc 
      ctctttgaa gtggacgat tttctggaa tccttgacc 
      acgctgtcc aaggaagaa gatcagtgc aaccgcttc 
      aaaactttc atcaaaaag tgcacagcg aatgacttg 
      aaagtggtc attcgcttg atcaaacac gacttaagg 
      atgagcgca ggagcgaaa catgtgttc aacggactg 
      catccggag gcctattcg gtcttccag acgtgcaaa 
      gacatcgcc actgtgata agcacttgt caggggcgc 
      aaagaaccc ccgaagaaa aggcggaaa atttccgaa 
      gagaaacgc aaatcgatt ccgaattat ttcgaaaat 
      gttaggttg tttttggag gagggggtg aaagagaga 
      cacgcccag tgggtgaaa tattttatt gcctacggt 
      gggtcggtt ctcaccaac gacaaaccc gaagaagcg 
      actcatgtt ttgcatttg aaagacact atttcacag 
      ccgacgatt cactgtgcc aacagcgca aaacatgtt 
      attgtggag tggattaaa gatacagta gcagaagag 
      aatttgcag gattatagg cggtataca gtccggtgg 
      gatccttgc cataaggat taa
    mRNA sequence
    • >TC004521 RA
      ggcaaagac gtaaacttt attgtttcg ctaaaataa 
      aataaataa ataggtttg tacagtggg taacgtgta 
      accgttgaa ctgatcagc tgttagtgt tgttgccag 
      tttttgagg aaaaagaaa gatggagga gaagccctt 
      cgccgtgga ggtggccag gcaagggcg cgccctttg 
      caaaaaatg ccgccagaa gtgcgttca aggcgattt 
      gcgcattgc taaattaat cccgaaccc tctgggttc 
      cggtaaaat gaaaacttg gtaccacgt aagctgtat 
      ctttgaaac atttttaaa acagagaca aacaactaa 
      aagaatcga gggccccct ggacatcga cggctggga 
      gtcactcga ccccgacga taaagtgaa tatcttgga 
      aaagatctc gcaatgtaa ccgggaatt tgaagccaa 
      gtttagtaa aaaaagacc gaaaaaggg gaccaagcc 
      gcagccgca gcttgtcgt cgacgacaa tttgagtaa 
      tttatctga taattgcac gactgagga ggttcacaa 
      ggataattg tttccgcga gtttcgcag gatcgtgtc 
      gaagctccc gaaggtgtc aagttccaa cgaaaaaat 
      cagcatttt ccgcgagat tttcacgaa aggttgcga 
      cgggagcgg ttttaaggg cgatgtcgt tctctggtg 
      caaactgct cctccctgc ggtgaacaa acgcgtgtt 
      tcacttgca gaccaagca actgattaa aatatttag 
      caagatttt cgcggttag tgaagaaac aatgactgc 
      tcacttgga acagggcga tgtgggcca gaccatcca 
      acatttctt cgaagaaaa cgccgagtg taaaccggc 
      gaaaaaaag caccttgag cctctttga agtggacga 
      ttttctgga atccttgac cacgctgtc caaggaaga 
      agatcagtg caaccgctt caaaacttt catcaaaaa 
      gtgcacagc gaatgactt gaaagtggt cattcgctt 
      gatcaaaca cgacttaag gatgagcgc aggagcgaa 
      acatgtgtt caacggact gcatccgga ggcctattc 
      ggtcttcca gacgtgcaa agacatcgc cactgtgat 
      aagcacttg tcaggggcg caaagaacc cccgaagaa 
      aaggcggaa aatttccga agagaaacg caaatcgat 
      tccgaatta tttcgaaaa tgttaggtt gtttttgga 
      ggagggggt gaaagagag acacgccca gtgggtgaa 
      atattttat tgcctacgg tgggtcggt tctcaccaa 
      cgacaaacc cgaagaagc gactcatgt tttgcattt 
      gaaagacac tatttcaca gccgacgat tcactgtgc 
      caacagcgc aaaacatgt tattgtgga gtggattaa 
      agatacagt agcagaaga gaatttgca ggattatag 
      gcggtatac agtccggtg ggatccttg ccataagga 
      ttaaacaat ttgttatta ttactcttt tatcaattt 
      ttcagaaga aaataaata caaaatacg aat
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY