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Gene information
General information
TC008019 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_007419.2: 3995076..3997490
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
    		TC008019
    iBeetle screen iB_01303

    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: 2 (3 dpi)
    Days post injection

    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature fluorescence decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
     head capsule mostly not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    head capsule complete not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    thorax cuticle dorsal irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    prothorax not present - Penetrance (percentage of animals/eggs on slide): less than 30%
    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - 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: 0.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 40.0% (includes death as larva, prepupa, pupa, adult)

    pupal molt delayed (still prepupa) - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection

    adult lethality - Number of animals showing the phenotype: 1 (16 dpi)
    Days post injection

    eclosion delayed (still pupa) - Number of animals showing the phenotype: 3 (16 dpi)
    Days post injection

    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 3 (19 dpi)
    Days post injection

    Fertility
    egg production: number <50% of expected -
    previtellogenic egg chamber not present - Number of animals showing the phenotype: 3

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC008019 RA
      MLNFQLTTE KDRREAALI DRRRAIEEE RKKRIFNPR 
QRLYGIDYE TLQRQIEEK RQREAQQAR IDRIFDEQR 
KKNDEMALA LEKKEKQER MKLEQEINY FRMCCQKPE 
DRREFDLND PNLLKKARP ARTADDDPK CGPSSAQKF 
EGEDLVSTE RAKVQRDQV KSWLDQQIM EKEAADKER 
KAAEEAYRE AIKARDQRA LQLDKMEKE CRKRLEEAC 
LRFNRALAD ERKMDEMKR LKREHDDCL AEMYNFMTS 
DLLTENPDV SQSNLGPNR RIGYLYKGM TDEEKKQVR 
EYQLAQIEE AKKRKELEK RMNREYDDY INGTQRTIS 
LLDKEEARK QREMVKSLA EENRNRL
    CDS sequence
    • >TC008019 RA
      atgttgaac ttccaatta acaacagag aaagaccgg 
cgagaagcc gccctcatc gaccggcgg cgcgccatc 
gaagaagag cgcaagaaa cgcattttc aaccctcgt 
cagcgctta tacggaata gattatgaa actttacaa 
agacagatc gaagagaaa aggcagcgg gaggcccaa 
caggcccgc atcgaccga atattcgat gagcagcgc 
aagaaaaac gacgaaatg gccctagcg ctcgagaag 
aaggagaaa caggaacgt atgaaactc gagcaagaa 
ataaattac ttccggatg tgctgccag aagccggaa 
gatcggcgc gaattcgat ttgaacgac ccgaacctt 
ttgaagaag gctaggccg gcgcgcacc gccgacgac 
gaccccaaa tgcggaccc tcctcagcc caaaagttt 
gaaggtgag gacctggtc agcaccgaa agggccaaa 
gttcagcga gatcaagtc aaatcgtgg ctcgaccaa 
cagataatg gagaaagaa gccgccgat aaggagcgc 
aaagcggcg gaagaggcc tacagagag gcgatcaag 
gcacgagat caaagagcc ctccagcta gataaaatg 
gagaaagag tgccggaag cggctcgaa gaggcgtgt 
ctcaggttc aacagggcg ttggccgat gaacgcaaa 
atggacgaa atgaagagg ctaaagagg gagcatgat 
gattgtctc gccgaaatg tacaatttt atgaccagc 
gatttgctg acggaaaat ccagacgtg tcccaaagt 
aatttaggg ccgaatcgc agaattgga tacttgtat 
aaggggatg accgatgag gagaaaaag caagtcagg 
gagtaccaa ttggcacaa attgaagag gcaaagaag 
cgcaaagag ttggaaaag cggatgaat cgggaatac 
gacgattac attaacgga acgcaacgc acaatatca 
ttactggac aaagaagaa gcccggaaa cagagggaa 
atggtgaaa agtttagct gaggaaaac cgaaatcgt 
ttataa
    mRNA sequence
    • >TC008019 RA
      ttatttaca aacgcctca aactctgac aaaatctgt 
gacactttt cccaaaaat gttgaactt ccaattaac 
aacagagaa agaccggcg agaagccgc cctcatcga 
ccggcggcg cgccatcga agaagagcg caagaaacg 
cattttcaa ccctcgtca gcgcttata cggaataga 
ttatgaaac tttacaaag acagatcga agagaaaag 
gcagcggga ggcccaaca ggcccgcat cgaccgaat 
attcgatga gcagcgcaa gaaaaacga cgaaatggc 
cctagcgct cgagaagaa ggagaaaca ggaacgtat 
gaaactcga gcaagaaat aaattactt ccggatgtg 
ctgccagaa gccggaaga tcggcgcga attcgattt 
gaacgaccc gaacctttt gaagaaggc taggccggc 
gcgcaccgc cgacgacga ccccaaatg cggaccctc 
ctcagccca aaagtttga aggtgagga cctggtcag 
caccgaaag ggccaaagt tcagcgaga tcaagtcaa 
atcgtggct cgaccaaca gataatgga gaaagaagc 
cgccgataa ggagcgcaa agcggcgga agaggccta 
cagagaggc gatcaaggc acgagatca aagagccct 
ccagctaga taaaatgga gaaagagtg ccggaagcg 
gctcgaaga ggcgtgtct caggttcaa cagggcgtt 
ggccgatga acgcaaaat ggacgaaat gaagaggct 
aaagaggga gcatgatga ttgtctcgc cgaaatgta 
caattttat gaccagcga tttgctgac ggaaaatcc 
agacgtgtc ccaaagtaa tttagggcc gaatcgcag 
aattggata cttgtataa ggggatgac cgatgagga 
gaaaaagca agtcaggga gtaccaatt ggcacaaat 
tgaagaggc aaagaagcg caaagagtt ggaaaagcg 
gatgaatcg ggaatacga cgattacat taacggaac 
gcaacgcac aatatcatt actggacaa agaagaagc 
ccggaaaca gagggaaat ggtgaaaag tttagctga 
ggaaaaccg aaatcgttt ataaaaaca cgccaacgt 
cggaattct acgaccagt ttaataaat caacaag
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY