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Gene information
General information
TC033320
Former TC numbers

If one gene is splitted or genes are merged, the original TC numbers are listed. One listed former TC number indicates the previous gene has be splitted, serveral former TC number represents a merged gene.

TC015307
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_007421.3: 661647..666169
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
    TC033320
    iBeetle screen iB_10156

    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: 0.0% (includes death as pupa, adult)

    Adult morphology
    abdominal odoriferous glands anterior content darker - Number of animals showing the phenotype: 5 (13 dpi)
    Days post injection



    abdominal odoriferous glands content not present - Number of animals showing the phenotype: 2 (13 dpi)
    Days post injection



    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 embryo/egg with embryonic tissue potentially present, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): 50% - 80%

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

    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: < 20
    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
    • >TC033320 RA
      MARIRWLTR TSRIVALCS AANFINSAD RVIMPIAIV 
      PMTDIFQWS LHAQGWILS AFAFGYFTS QILGSFWAS 
      KLGAKNVLI CSVLLWSIA TFITPFLAH SITALVICR 
      IILGFAEGL GLPTIFHLF AHSIPVEER SRAFSYLLA 
      SGSVGQTVA SVLCPHLSW EACFYWFGS FGFVWVVLW 
      YFTYPEDAF SAEETIPLH MPKVISRNI RWADFILSK 
      PLWAIYLAH FAMNWSSYI IMQWLPTYL ARYLGGNAH 
      SLSLTAVPY VVNSVFGVI SGHVADGLL LKGWSVLNV 
      RRLMTSVGL VGPALFLAI FCVVDHLPF AVILVSISM 
      GLSASNSAG HLSNHVDVA PNYAGTTFA ISNTIATIP 
      GILCGPVTA SLVTASDGK WMSVFLGAT FVNLMGAFV 
      YFTNSVATQ VL
    CDS sequence
    • >TC033320 RA
      atggctagg attcgctgg ttgacgagg acctcgcgc 
      attgtggcc ctctgcagt gctgccaat tttataaat 
      agcgcggat agggtgatt atgcccatt gctattgta 
      ccaatgacc gatattttt caatggagt ttacacgcc 
      caaggctgg attttgtcg gcatttgcc ttcggatat 
      ttcactagt caaatatta ggcagtttt tgggcgagt 
      aaattgggc gctaaaaac gtcctaatc tgcagcgtt 
      ctcttatgg agcatcgcc acattcatc acccccttt 
      ctggcccac agcattaca gctttagtc atttgccga 
      ataattctc ggttttgct gaaggacta ggtcttcca 
      acaattttc catctattc gcccacagc ataccagtt 
      gaggaacgt tcgcgtgct ttcagttat ctgctagct 
      tcaggctca gttggccaa actgtggct tcagtcctg 
      tgcccccac ttatcctgg gaggcttgt ttctactgg 
      ttcggctca tttggcttt gtttgggtg gttttgtgg 
      tactttacc tacccagag gacgctttc agtgccgag 
      gaaaccata ccattgcat atgcccaaa gtgatatca 
      cgtaatatc agatgggcc gattttatt ttaagcaaa 
      ccattatgg gccatttac ttggcgcat tttgccatg 
      aactggtcg tcttacatc ataatgcag tggctgccc 
      acttatctg gcgcgctat ttggggggc aatgcccac 
      agtttgagc ctaacggca gtgccgtac gttgttaat 
      tcggtcttt ggggtgata agtgggcac gtggctgat 
      gggctacta ctcaaaggt tggtcagtg ctgaatgtc 
      cgtcggttg atgaccagt gtggggctc gtggggccc 
      gcgctcttt ttggccatt ttctgtgtc gtggaccat 
      ttgccgttt gccgtcatt ctggtgagc atctcgatg 
      gggctgtcg gccagcaat tcggccgga catttgagt 
      aatcacgtg gatgtggcc cccaattat gccgggacc 
      acgtttgcg attagtaat acgattgcc acgattccg 
      ggcattttg tgcgggcca gtgactgcg tccctggtc 
      acggcctca gatggcaag tggatgagc gtctttctg 
      ggggccacg tttgtcaat ctcatgggg gcgttcgtc 
      tatttcacg aatagcgtg gccacgcag gtcttgtga
    mRNA sequence
    • >TC033320 RA
      acattgttg ttgacatgg ccatgccaa acgtcaagt 
      ttgcgttaa ttttgcaaa aaaacgcct aatttgaaa 
      taaaaacct cctaatggt agacttaag ttaggtttt 
      atttacttt ggaatggtt taagtgcgg tttatgaca 
      agccttaaa atggctagg attcgctgg ttgacgagg 
      acctcgcgc attgtggcc ctctgcagt gctgccaat 
      tttataaat agcgcggat agggtgatt atgcccatt 
      gctattgta ccaatgacc gatattttt caatggagt 
      ttacacgcc caaggctgg attttgtcg gcatttgcc 
      ttcggatat ttcactagt caaatatta ggcagtttt 
      tgggcgagt aaattgggc gctaaaaac gtcctaatc 
      tgcagcgtt ctcttatgg agcatcgcc acattcatc 
      acccccttt ctggcccac agcattaca gctttagtc 
      atttgccga ataattctc ggttttgct gaaggacta 
      ggtcttcca acaattttc catctattc gcccacagc 
      ataccagtt gaggaacgt tcgcgtgct ttcagttat 
      ctgctagct tcaggctca gttggccaa actgtggct 
      tcagtcctg tgcccccac ttatcctgg gaggcttgt 
      ttctactgg ttcggctca tttggcttt gtttgggtg 
      gttttgtgg tactttacc tacccagag gacgctttc 
      agtgccgag gaaaccata ccattgcat atgcccaaa 
      gtgatatca cgtaatatc agatgggcc gattttatt 
      ttaagcaaa ccattatgg gccatttac ttggcgcat 
      tttgccatg aactggtcg tcttacatc ataatgcag 
      tggctgccc acttatctg gcgcgctat ttggggggc 
      aatgcccac agtttgagc ctaacggca gtgccgtac 
      gttgttaat tcggtcttt ggggtgata agtgggcac 
      gtggctgat gggctacta ctcaaaggt tggtcagtg 
      ctgaatgtc cgtcggttg atgaccagt gtggggctc 
      gtggggccc gcgctcttt ttggccatt ttctgtgtc 
      gtggaccat ttgccgttt gccgtcatt ctggtgagc 
      atctcgatg gggctgtcg gccagcaat tcggccgga 
      catttgagt aatcacgtg gatgtggcc cccaattat 
      gccgggacc acgtttgcg attagtaat acgattgcc 
      acgattccg ggcattttg tgcgggcca gtgactgcg 
      tccctggtc acggcctca gatggcaag tggatgagc 
      gtctttctg ggggccacg tttgtcaat ctcatgggg 
      gcgttcgtc tatttcacg aatagcgtg gccacgcag 
      gtcttgtga atattgtat atatttgtg tagattaat 
      aaaagaata ccaaactt
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY