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Gene information
General information
TC003966 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: 29938046..29939611
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
    		TC003966
    iBeetle screen iB_07092

    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: 30.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 3 (3 dpi)
    Days post injection

    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): more than 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    remarks: muscle pattern potentially obscured by segmentation defect,
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
     embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    cuticle remnants with some identifiable segments - Penetrance (percentage of animals/eggs on slide): less than 30%

    larva everted (inside-out) - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    head capsule size decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    thorax segment number decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    thorax number decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    abdomen segment number decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    abdomen number decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    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
    • >TC003966 RA
      MTQALLNRC RMHTLSLFP LLKPPKMPT TVTQFHTTP 
NPHRKSLYK TESKKIRFF VVHRYLEYL KNYDKVLER 
SFPGAMRVY RVFTVGIKD FAQDLKDYF RIVRILNSP 
TKDKFKSLT RREIELYHQ MPKDMRKVA PVLLISALP 
FANYVIFPL AYLFPRIFL SRHFWNLQQ RAEFSIYYL 
RGRLVNNRP LFRCVQGEL DKIRGHRLY YKWAEVLGM 
MGSGVQPKV EQILQCKDL FTCEPYHLF YLKGKHVKH 
LLKIHDLHM GWFRRSRLI ERAVILQEM DKAIVREGG 
VHNMPQDAL KHACYIRGL NPMNMRNED MINWLNNWI 
KISSVVDRE SWSLLLHCP ILLAYNEPS NWTLIYKPK
    CDS sequence
    • >TC003966 RA
      atgactcaa gctctcctc aaccggtgc agaatgcac 
actttgagc cttttcccg ctactaaaa cccccgaaa 
atgccaaca actgtcacg caatttcac acgacacct 
aacccccat cggaagtcc ctctacaag actgagtcg 
aagaaaatc cgctttttc gtcgtccac cgttacctc 
gaatatttg aaaaactat gacaaggtc ttggagcgc 
agctttccg ggggccatg cgggtctac cgcgttttc 
acagtcggt attaaagac tttgcccag gacttgaag 
gactacttc cgaatagtg cgaattcta aactcccca 
accaaagac aagtttaaa agtttgacg cggcgcgag 
atcgagctc taccaccag atgcccaaa gacatgcgg 
aaggtggcc cccgtgttg ctgatttca gccctacct 
ttcgccaat tacgttatt ttcccgttg gcttatttg 
ttcccgagg attttcctc tcgcgtcat ttctggaat 
ttgcaacaa agggccgag tttagtatt tattatctg 
agggggcgt ctggtgaac aatcggccg ctttttcgg 
tgcgttcaa ggcgagttg gataagata agggggcac 
aggttgtac tacaaatgg gccgaagtt ttgggcatg 
atgggcagt ggcgtccag cccaaggtt gagcagatt 
ctacagtgc aaggatttg tttacgtgc gagccgtac 
catctgttt tatttgaag ggaaaacat gtgaaacac 
ttgttgaaa atccatgac ctccacatg gggtggttt 
cggcgctcg cgcctcatt gaacgagct gtgatactg 
caagaaatg gacaaggcc atagtcagg gaaggaggc 
gtccacaac atgccccaa gacgcctta aaacacgct 
tgttacata agggggctc aacccgatg aatatgcga 
aacgaagac atgataaac tggctaaac aactggatc 
aaaatttcc agtgttgtt gacagggaa agttggtca 
cttctgctg cactgtccc attttactg gcttacaac 
gagccctca aactggaca ctgatttac aaacccaag 
tga
    mRNA sequence
    • >TC003966 RA
      tgtcaaaaa tatctaacc ttcaaacga tagcattac 
gaaaatagt tttaaatcc gaataacca gtgtgaaaa 
cgatcaata tgggcaaaa tcgaggacg acaaataaa 
taccaagcg ttcgaggtg cccaaatcg ataaatcac 
aatgactca agctctcct caaccggtg cagaatgca 
cactttgag ccttttccc gctactaaa acccccgaa 
aatgccaac aactgtcac gcaatttca cacgacacc 
taaccccca tcggaagtc cctctacaa gactgagtc 
gaagaaaat ccgcttttt cgtcgtcca ccgttacct 
cgaatattt gaaaaacta tgacaaggt cttggagcg 
cagctttcc gggggccat gcgggtcta ccgcgtttt 
cacagtcgg tattaaaga ctttgccca ggacttgaa 
ggactactt ccgaatagt gcgaattct aaactcccc 
aaccaaaga caagtttaa aagtttgac gcggcgcga 
gatcgagct ctaccacca gatgcccaa agacatgcg 
gaaggtggc ccccgtgtt gctgatttc agccctacc 
tttcgccaa ttacgttat tttcccgtt ggcttattt 
gttcccgag gattttcct ctcgcgtca tttctggaa 
tttgcaaca aagggccga gtttagtat ttattatct 
gagggggcg tctggtgaa caatcggcc gctttttcg 
gtgcgttca aggcgagtt ggataagat aagggggca 
caggttgta ctacaaatg ggccgaagt tttgggcat 
gatgggcag tggcgtcca gcccaaggt tgagcagat 
tctacagtg caaggattt gtttacgtg cgagccgta 
ccatctgtt ttatttgaa gggaaaaca tgtgaaaca 
cttgttgaa aatccatga cctccacat ggggtggtt 
tcggcgctc gcgcctcat tgaacgagc tgtgatact 
gcaagaaat ggacaaggc catagtcag ggaaggagg 
cgtccacaa catgcccca agacgcctt aaaacacgc 
ttgttacat aagggggct caacccgat gaatatgcg 
aaacgaaga catgataaa ctggctaaa caactggat 
caaaatttc cagtgttgt tgacaggga aagttggtc 
acttctgct gcactgtcc cattttact ggcttacaa 
cgagccctc aaactggac actgattta caaacccaa 
gtgattatg ttactagtt ataataatt tatttttgt 
gataaatga taatgaacc aaaagttta attttgtat 
taaattagt aacaacaag
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY