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Gene information
General information
TC005849 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_007423.3: 6625244..6632267
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
    TC005849
    iBeetle screen iB_04022

    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: 10.0% (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


    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): more than 80%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - 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 total percentage of affected eggs/embryos/larvae on slide - 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: 20.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 30.0% (includes death as larva, prepupa, pupa, adult)

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


    adult lethality - Number of animals showing the phenotype: 1 (22 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 2 (41 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC005849 RA
      MEISFEGKR ALVTGASRG IGRAIVGQL AKCGAQVVA 
      VGRSKADLE ALKAETGVE TLILDISDW DKTEKVLSQ 
      LDSIDLLVN NAGEGMIKP LIDLEEGDI DRIFSLNLK 
      GLINVTRLI AKNMIRKKS GAIVNISSQ ASLAGLLNH 
      TVYSASKGG VDAFTRACA LELGPHNIR VNSVNPTVI 
      MTEMGRKWW SEPSRAGTM LHKIPLGRF GEVQEVVDA 
      VLFLLSDKA AMITGTCLP IDGGFVSS
    CDS sequence
    • >TC005849 RA
      atggagatc tcctttgag ggtaaaagg gcgctagtt 
      acgggggct tcgaggggg atagggagg gctattgtg 
      ggccagttg gccaaatgt ggggcccaa gttgttgcc 
      gtgggacgc tccaaagct gacttagag gcactgaaa 
      gctgaaaca ggggttgaa accctgatt ttggacatc 
      tcggactgg gacaagacc gaaaaagtt ttaagccaa 
      ctcgattcg attgattta ctggtcaat aacgccggc 
      gaagggatg atcaaacct ttgattgat ttggaagaa 
      ggggatatt gaccgaatt ttttccttg aatttgaag 
      ggcctaatt aatgttact cgcttaata gccaaaaac 
      atgattagg aaaaaatca ggtgcgatt gtaaatatc 
      tcatctcaa gcctctttg gctggtctt ttgaaccac 
      actgtttat tcggcgagt aaaggtgga gtcgacgct 
      tttaccaga gcttgtgcc cttgagttg ggtcctcac 
      aacattagg gttaattcg gtcaatccc actgtgatt 
      atgactgaa atgggccga aaatggtgg agcgaaccg 
      tcacgtgct gggacaatg ctacacaaa ataccattg 
      ggacggttt ggggaagta caggaagtg gtcgatgcc 
      gttttgttt ctcctgagc gacaaggct gccatgatt 
      acggggacg tgtctcccc attgacggc ggttttgta 
      tcgtcttaa
    mRNA sequence
    • >TC005849 RA
      tttttcgcc cgtgatagg ttggtaaga gtaaaaatt 
      ataattatt tatttcaaa tgtataatt attcccata 
      tttaggtag gtaactaat agaaataac caacaatat 
      tttcctgta aatattttt tactgtgca gggattggt 
      cgagacatt gtgaaacaa ttggcgaag ggcggggcc 
      aaggtggtg gccgttgcc cgaaacaag gaactattg 
      gagagcctc aaagaggag gttccctca atcgaaatc 
      attccttgc gacctctct gattggaac gcgacccaa 
      aatgcctta tcccacctt tctcccgac ctcctggtc 
      aacaacgca ggccttgca attctaaaa cccctaacc 
      cgggttagc gaatcggac gttgaccaa cttttcgcc 
      ataaatgtc aaagccttg atccaggtt actaaacta 
      gtaaccgag aagttactc caaaggggg gtttccggt 
      tccattgtg aatatctcg tcccaagcc tcaatggct 
      gggcttttg aaccactcc gtctactgc agtaccaag 
      ggggctgtc gatgcgttc actcgagcc gcagccctg 
      gagcttggc ccccataat atcagagtt aattgtgtt 
      aacccaact gttataatg acggcaatg gggcgcttg 
      ggttggtcg gagccgaaa gtggccgaa cccatgttg 
      cagaaaata ccgttgagg aggtttgga gaggttcaa 
      gaggttgtt gatgcggtt ttgttttta ttgagtgat 
      aaggcgagc atgattacg gggacttgt cttcctgta 
      gatggcggt tatctcgct gtttaataa taataatta 
      ttaataaag atgttattc agtcgtaaa agttgtttg 
      attcgttca tcactccaa aatggagat ctcctttga 
      gggtaaaag ggcgctagt tacgggggc ttcgagggg 
      gatagggag ggctattgt gggccagtt ggccaaatg 
      tggggccca agttgttgc cgtgggacg ctccaaagc 
      tgacttaga ggcactgaa agctgaaac aggggttga 
      aaccctgat tttggacat ctcggactg ggacaagac 
      cgaaaaagt tttaagcca actcgattc gattgattt 
      actggtcaa taacgccgg cgaagggat gatcaaacc 
      tttgattga tttggaaga aggggatat tgaccgaat 
      tttttcctt gaatttgaa gggcctaat taatgttac 
      tcgcttaat agccaaaaa catgattag gaaaaaatc 
      aggtgcgat tgtaaatat ctcatctca agcctcttt 
      ggctggtct tttgaacca cactgttta ttcggcgag 
      taaaggtgg agtcgacgc ttttaccag agcttgtgc 
      ccttgagtt gggtcctca caacattag ggttaattc 
      ggtcaatcc cactgtgat tatgactga aatgggccg 
      aaaatggtg gagcgaacc gtcacgtgc tgggacaat 
      gctacacaa aataccatt gggacggtt tggggaagt 
      acaggaagt ggtcgatgc cgttttgtt tctcctgag 
      cgacaaggc tgccatgat tacggggac gtgtctccc 
      cattgacgg cggttttgt atcgtctta attttagtt 
      gaattaatt gtcaaaaat ttattgttg gaaaaagag 
      agattaaag ataggacca attg
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY