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Gene information
General information
TC003347 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_007418.3: 1721103..1722548
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
    		TC003347
    iBeetle screen iB_00559

    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)

    adult lethality - Number of animals showing the phenotype: 2 (9 dpi)
    Days post injection

    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: < 50 embryo/egg not developed/not fertilized - 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
     egg without cuticle 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: 0.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 20.0% (includes death as larva, prepupa, pupa, adult)

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

    eclosion delayed (still pupa) - Number of animals showing the phenotype: 1 (16 dpi)
    Days post injection

    adult lethality - Number of animals showing the phenotype: 1 (19 dpi)
    Days post injection

    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (19 dpi)
    Days post injection

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC003347 RA
      MRPKLYMSE ICPSARAVV LTAKVLELT 
LELKEVSCD KKLNQQFSI PTLEDSGYV 
IWDSHAIIA FLVGKYGKD DSLYPRDNP 
RRAIIDERL RFDSGVVSF FTKTILNSI 
PYEDNEKAV NEIYSLVEE FFDGNNPWI 
AGDALSIAD LSLIPSITS LDVVVPIDP 
KRFPKLARW VKRAETMPF FEANKTGLC 
KLRRILNRC
    CDS sequence
    • >TC003347 RA
      atgcgcccc aagttgtac atgagcgag atatgtccc 
tcagctaga gccgtcgtg ttgacagcc aaggtcctc 
gaactcacc ctcgagctg aaggaggtt tcttgcgac 
aaaaagttg aatcaacaa ttctccatc cctacgctg 
gaggacagc ggctacgtg atttgggac agccatgcc 
attattgcc ttcctggtc gggaagtac ggcaaagac 
gactccttg taccccaga gacaacccc cggagggcc 
attatagac gaacgattg cggtttgac tcgggggtc 
gtgtctttt ttcaccaaa acgattctg aactcgata 
ccatacgaa gataatgaa aaagccgtc aatgaaatt 
tattccttg gttgaggaa ttcttcgat ggtaacaac 
ccatggatc gctggggat gctttaagc atcgcagac 
ttaagtcta attccttcg atcacgtcg ctggatgtt 
gtggtccct attgacccc aagcggttc cctaagtta 
gcaagatgg gtgaaaaga gccgaaaca atgcccttt 
tttgaggca aataagacc gggctctgt aaactccga 
aggattcta aaccggtgt taa
    mRNA sequence
    • >TC003347 RA
      gcagacagc gataacaaa aaaccgctc ctctcctca 
gttcagcca cactcgcaa taacaaaaa cattcaaca 
gctctgttt atccccagt tcaaaatgc gccccaagt 
tgtacatga gcgagatat gtccctcag ctagagccg 
tcgtgttga cagccaagg tcctcgaac tcaccctcg 
agctgaagg aggtttctt gcgacaaaa agttgaatc 
aacaattct ccatcccta cgctggagg acagcggct 
acgtgattt gggacagcc atgccatta ttgccttcc 
tggtcggga agtacggca aagacgact ccttgtacc 
ccagagaca acccccgga gggccatta tagacgaac 
gattgcggt ttgactcgg gggtcgtgt cttttttca 
ccaaaacga ttctgaact cgataccat acgaagata 
atgaaaaag ccgtcaatg aaatttatt ccttggttg 
aggaattct tcgatggta acaacccat ggatcgctg 
gggatgctt taagcatcg cagacttaa gtctaattc 
cttcgatca cgtcgctgg atgttgtgg tccctattg 
accccaagc ggttcccta agttagcaa gatgggtga 
aaagagccg aaacaatgc ccttttttg aggcaaata 
agaccgggc tctgtaaac tccgaagga ttctaaacc 
ggtgttaac gctaataat aatacaact tttattgta 
actttaagt gattaattg tgatatgag gctattagt 
tttagatat ggatttttt atttgatgt aatattaat 
aaatgaagc gataaaaag acgtgcgat gccaggtgt 
aattcgtgg ttaatttaa cttgcaggc tcgcttttt 
tgtccgaga tatgttata tcattgttt ggacaatga 
ccaagtagc acaacatta gtttctgat atggcgtgc 
gttaatgcg atctgttta tcgtttgta attacatca 
cgtggagat gtgcaacga aatccgggt gataagagt 
tttatttaa gtcatctat tattaagta cttaatcta 
gaaagtaca cgtttgacc ccaactaaa cttaaattt 
tatcagctc gtttgtgaa cccaaacac ttaggattt 
ttcattttg gcttatctt ttatcagaa agtgaatta 
tgtggtttt attagatac ttgatcgaa ctcatcaga 
caaaattaa tactacatg gttttactt ttatattat 
acaatttgt ttttacgac caataaaac actctccag 
ttt
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