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Gene information
General information
TC002096 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.

NW_015452015.1: 7057..8883
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
    TC002096
    iBeetle screen iB_03303

    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


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 embryo/egg with embryonic tissue, no cuticle, but muscle fluorescence - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    larval musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    remarks: muscle pattern potentially obscured by segmentation defect,
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 50
    head appendages rounded - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    head appendages size decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    abdomen segment number decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae - 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: % (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
    • >TC002096 RA
      MIQVTWKIK HLLVLLASL SILLLVFIK YSRQRSAEN 
      VHNLHLYQD ELEYNKHLL QEASERIQD LENKILALE 
      ARVPKTYPE VKFLNYLSR KRILITGGA GFVGSHLVD 
      RLMLQGHEV IVADNFFTG RKRNVEHWI GHENFELIH 
      HDIVNPLFI EVDEIYHLA SPASPPHYM YNPVKTIKT 
      NTLGTINML GLARRLNAK ILIASTSEV YGDPDIHPQ 
      PETYWGHVN PIGPRACYD EGKRVSETL TYAYAKQEN 
      MQVRVARIF NTYGPRMHM NDGRVVSNF ILQALQNDV 
      ITIYGSGQQ TRSFQYISD LVDGLVALM NSNYTLPVN 
      LGNPVEHSI NEFASIIKD LVGGRSKIN HLAEVEDDP 
      QRRRPDITR AKKYLNWEP KVDLNTGLQ KTVDYFRQE 
      LHRFKHSQR NKFLPKL
    CDS sequence
    • >TC002096 RA
      atgattcag gttacttgg aaaatcaaa cacttgctt 
      gtactacta gcgtcctta tcgattttg ttgttggtt 
      tttattaaa tacagccgg caaaggtcg gccgaaaat 
      gtgcacaat ttgcacctg tatcaggac gagcttgag 
      tacaacaag caccttctg caagaagcg tccgagcga 
      attcaagac ttggaaaat aaaatccta gctcttgag 
      gcaagggtg cccaagacg taccccgaa gttaagttc 
      ctaaactac ctctcgcga aaacgtatt ttgataact 
      ggcggtgcc ggtttcgtg gggtcacac ctagttgat 
      agattaatg ctgcaaggt catgaagtc atcgtcgct 
      gataatttt ttcacgggg cggaagcgc aacgtcgag 
      cactggata ggtcacgag aatttcgaa ttaattcac 
      cacgacata gtcaatcct ttatttatt gaagtcgac 
      gagatctac catctcgca agccccgct agtcccccc 
      cattacatg tacaacccg gtcaaaacc atcaaaact 
      aacacttta ggtacaatt aacatgcta ggccttgcc 
      agaagactg aacgccaaa attctaatc gccagcact 
      tcggaagtg tacggagac cccgacatc cacccccag 
      cccgagacg tactgggga catgtcaat cccataggc 
      cctcgcgcg tgttacgac gaaggcaaa cgagtctca 
      gagacttta acttatgct tatgccaag caggaaaac 
      atgcaggtg cgggtggcc cgcattttc aacacttac 
      ggcccccgt atgcacatg aatgacggc agagtcgtc 
      tctaatttt atcctccag gcgctgcaa aacgacgta 
      atcacgatt tacggctca gggcagcaa acacgctcg 
      ttccagtac atttccgac ctggtcgat gggctggta 
      gcgctgatg aacagcaac tatacgctc ccggtgaac 
      ttggggaat cccgtggag cattcaatt aacgaattt 
      gcctcaata attaaagac ctggtgggc ggccgcagc 
      aaaatcaat catttggcg gaagtggag gacgatccg 
      cagcgccgc aggccggac atcacccgc gcgaaaaaa 
      tacctaaac tgggagccg aaggtggac ctcaacacc 
      ggcctccaa aagactgtc gactacttc cgccaggag 
      ctgcaccgg tttaagcac tcacagaga aacaagttt 
      ttaccgaaa ttgtga
    mRNA sequence
    • >TC002096 RA
      caattaggc gtttttaat ggataaaaa atgcgatat 
      cgttgcgta acatgattc aggttactt ggaaaatca 
      aacacttgc ttgtactac tagcgtcct tatcgattt 
      tgttgttgg tttttatta aatacagcc ggcaaaggt 
      cggccgaaa atgtgcaca atttgcacc tgtatcagg 
      acgagcttg agtacaaca agcaccttc tgcaagaag 
      cgtccgagc gaattcaag acttggaaa ataaaatcc 
      tagctcttg aggcaaggg tgcccaaga cgtaccccg 
      aagttaagt tcctaaact acctctcgc gaaaacgta 
      ttttgataa ctggcggtg ccggtttcg tggggtcac 
      acctagttg atagattaa tgctgcaag gtcatgaag 
      tcatcgtcg ctgataatt ttttcacgg ggcggaagc 
      gcaacgtcg agcactgga taggtcacg agaatttcg 
      aattaattc accacgaca tagtcaatc ctttattta 
      ttgaagtcg acgagatct accatctcg caagccccg 
      ctagtcccc cccattaca tgtacaacc cggtcaaaa 
      ccatcaaaa ctaacactt taggtacaa ttaacatgc 
      taggccttg ccagaagac tgaacgcca aaattctaa 
      tcgccagca cttcggaag tgtacggag accccgaca 
      tccaccccc agcccgaga cgtactggg gacatgtca 
      atcccatag gccctcgcg cgtgttacg acgaaggca 
      aacgagtct cagagactt taacttatg cttatgcca 
      agcaggaaa acatgcagg tgcgggtgg cccgcattt 
      tcaacactt acggccccc gtatgcaca tgaatgacg 
      gcagagtcg tctctaatt ttatcctcc aggcgctgc 
      aaaacgacg taatcacga tttacggct cagggcagc 
      aaacacgct cgttccagt acatttccg acctggtcg 
      atgggctgg tagcgctga tgaacagca actatacgc 
      tcccggtga acttgggga atcccgtgg agcattcaa 
      ttaacgaat ttgcctcaa taattaaag acctggtgg 
      gcggccgca gcaaaatca atcatttgg cggaagtgg 
      aggacgatc cgcagcgcc gcaggccgg acatcaccc 
      gcgcgaaaa aatacctaa actgggagc cgaaggtgg 
      acctcaaca ccggcctcc aaaagactg tcgactact 
      tccgccagg agctgcacc ggtttaagc actcacaga 
      gaaacaagt ttttaccga aattgtgat aattgtggt 
      tttgttgta aataaattt taaaatttt cg
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY