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Gene information
General information
TC008369 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: 9217981..9219456
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
    		TC008369
    iBeetle screen iB_08167

    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)

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

    Comment: larvae seem to die some time after hatching
    larval musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    remarks: muscle pattern potentially obscured by segmentation defect,
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
     larva partially everted (inside-out) - Penetrance (percentage of animals/eggs on slide): less than 30%

    head appendages shape irregular - Penetrance (percentage of animals/eggs on slide): more than 80%

    antenna proximal size decreased - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    legs size decreased - Penetrance (percentage of animals/eggs on slide): more than 80%

    legs thinner - 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
    • >TC008369 RA
      MLMQMIFSP GGAHQGPNK SEGLDDTGV 
APSPWANLL SVGLKILTA ILGGGAGGN 
SDGIDKVDN GSPMQGVLA AILSTVLGA 
RDPNQVNMM AKQAGEFIN IVVNLLDAL 
KTSFSHRSM AARNIGKKD SISDAAVAG 
IAMMKGYVR SLSTADSKC MQKYMCDAN 
SECSTDVGQ SSLFCHLGS YAASFILDR 
STSSTTFDL LYEAGRRGR SGDNCRQAY 
LECNEV
    CDS sequence
    • >TC008369 RA
      atgctcatg cagatgatc ttcagtccc ggcggcgcc 
caccaaggg cccaacaaa tccgaaggc ttggacgac 
acaggcgtt gcgccttct ccatgggcg aacttgctc 
tccgttggt ctgaaaatc ctcactgcg attcttgga 
ggcggcgcg ggcggaaac agcgacggc atcgacaaa 
gtcgacaac gggtcccct atgcagggc gttctggcg 
gcgattctt tcaacggtg cttggagct agggaccca 
aatcaggtc aacatgatg gctaagcaa gctggggag 
ttcatcaac atcgtggtg aacctcttg gacgctctg 
aagacatcc ttctcccac cggtccatg gcagccaga 
aacatcggc aagaaggac tcgattagc gacgccgcc 
gtcgccggg attgccatg atgaagggc tacgtgagg 
tcgctgtcc accgcagac agcaagtgc atgcagaag 
tacatgtgc gatgctaac agcgaatgc tcgacagat 
gttggccaa agctctctc ttctgtcac ctcggaagt 
tatgcggct agcttcatt ttggaccga tcgacgagc 
tccaccacc tttgattta ctctacgaa gctggaagg 
cgaggaaga tccggggac aactgccgt caagcatac 
ttggaatgc aacgaagtt tga
    mRNA sequence
    • >TC008369 RA
      ttctaggta tgctcatgc agatgatct tcagtcccg 
gcggcgccc accaagggc ccaacaaat ccgaaggct 
tggacgaca caggcgttg cgccttctc catgggcga 
acttgctct ccgttggtc tgaaaatcc tcactgcga 
ttcttggag gcggcgcgg gcggaaaca gcgacggca 
tcgacaaag tcgacaacg ggtccccta tgcagggcg 
ttctggcgg cgattcttt caacggtgc ttggagcta 
gggacccaa atcaggtca acatgatgg ctaagcaag 
ctggggagt tcatcaaca tcgtggtga acctcttgg 
acgctctga agacatcct tctcccacc ggtccatgg 
cagccagaa acatcggca agaaggact cgattagcg 
acgccgccg tcgccggga ttgccatga tgaagggct 
acgtgaggt cgctgtcca ccgcagaca gcaagtgca 
tgcagaagt acatgtgcg atgctaaca gcgaatgct 
cgacagatg ttggccaaa gctctctct tctgtcacc 
tcggaagtt atgcggcta gcttcattt tggaccgat 
cgacgagct ccaccacct ttgatttac tctacgaag 
ctggaaggc gaggaagat ccggggaca actgccgtc 
aagcatact tggaatgca acgaagttt gaaactcca 
ctttagaat cacacatat ccaaaatca gcaataatt 
tatttgtaa aatagtatt tattttttt cattgcatg 
gcacatgat tttaatcgg aaatcgatg gaaatcaag 
gactggtcc gagctttgt tagtggatg cggttattt 
tttatcatt tatgaatgc tgtatgaat ggatgaatg 
agcagcgaa tggggttca tgtacagta taaattata 
actcgatta caataaagt ttatttatt cattaaaac 
ctccattat tttttactt aaatagaac caaatttaa 
tcctaagga agtttcacc aaattgaga taacaaccc 
tgccttgca cagttatca caacatttt ggttaaact 
tagaaataa tccattgtt atgattatg ttaagttaa 
taaatgttt ctatgtaca aaatgagtt ttctttcaa 
ttttaccga aaaatgccg aaataaggc taactagct 
ttcaacata atccacagt ctgttggat tatagtagt 
aatacagcc cctatagga ca
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY