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Gene information
General information
TC003462 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: 5425849..5427014
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
    		TC003462
    Gene ontology

    GO terms for Tribolium

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    There are no GO terms for Tribolium at the moment. You can help the community by providing some GO terms for this gene.


    GO terms for the homologous Drosophila genes

    Assign a GO term to this Tribolium gene by filling in the fields. Repeat to add several GO terms. Search AmiGO for the correct GO ID. Use only the most specific term - use "graph views" to browse related terms ("child terms" are more specific; the more general "parent" terms will be automatically linked). Only information based on Tribolium data should be entered - do not define terms just based on Drosophila knowledge. We will review this information and submit the annotation to the Gene Ontology consortium.


    Gene
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    The Tribolium gene the GO term is assigned to.
    GO ID *
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    The ID of the GO term you assign to the gene. Please enter only the ID of the GO term with proceeding 'GO:'. This information is mandatory.
    Evidence
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    The evidence code describes with which method the function or location, expressed by the GO term, was show for this gene. Please refer to this page for a definition of the evidence code and a decision tree.
    Reference *
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    The reference where the function or location is described for the gene. Please enter a PubMed ID. This information is mandatory.
    Quotation
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    Name
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    Email
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    * : mandatory fields
    For help on how to submit larger datasets or non experimental data please contact us.
    iBeetle screen iB_09008

    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: 1 (3 dpi)
    Days post injection

    adult lethality - Number of animals showing the phenotype: 1 (9 dpi)
    Days post injection

    adult lethality - Number of animals showing the phenotype: 1 (11 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 strongly fragmented (cuticle crumbs) - 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
    • >TC003462 RA
      MSKPVVSKQ KSKKYYFKK TSNKRLSLD VNLRGFLCS 
CNNREKDCI KESYNLLNE YADKLYQES NEPEAEQDI 
DDSLAKEIS ELKQDKSEK RFQVIESGA KNFLFIRTS 
LENPVELAE AIIKDVDGS KTQRTKFLL RLIPVEITC 
KANVSDIVN AFVPLAQKH FVESPQTFC VIFNHRNNN 
VVSRDEVIK LIAAKVSEL RPDHKVDLK EAKVAIIVE 
VIKGFAFLS VIPDYLKHK KYNLLSLCS QEQVADCN
    CDS sequence
    • >TC003462 RA
      atgagtaag ccagttgtt tccaaacaa aaatccaaa 
aaatactac tttaaaaaa acttccaac aaaagactg 
agcttggac gtgaacttg cggggcttc ctctgcagt 
tgtaacaac agggaaaag gactgcatt aaggaatct 
tacaattta ctaaacgaa tatgctgac aaattgtac 
caagagagc aacgaacct gaagctgag caagacatt 
gacgactcg ttggctaaa gaaatatcc gaactgaag 
caagacaag agtgaaaaa cgattccag gttatagaa 
tctggtgcc aaaaatttc ctattcatt aggacgagt 
ttggagaat ccggttgag ctagcagag gcgattata 
aaagatgtt gatgggagt aagacccaa agaacgaaa 
tttttgctg agactgatt cctgtggaa attacgtgc 
aaggcgaat gtgagtgat attgtcaac gcatttgtg 
cccctcgcc cagaaacac tttgtggag tccccgcaa 
acgttttgt gtgatcttc aaccaccgg aataataat 
gttgtttcg cgggatgaa gtcataaaa ttaattgca 
gcgaaagtg tccgagctg aggcctgat cataaagtt 
gatcttaaa gaggcaaaa gtggctatt attgttgaa 
gtgataaag gggtttgcg tttttaagt gttatacct 
gactatctt aagcacaaa aagtataat ttattgtca 
ctttgtagt caagaacaa gtagctgat tgtaattaa
    mRNA sequence
    • >TC003462 RA
      ctaacctaa tttttaagt gcgtattta ttttaactt 
tttatgctt acaaataaa aaaatattg gattaatac 
gtgttatta acaaagaac agtggggtc gtagtcatg 
agtaagcca gttgtttcc aaacaaaaa tccaaaaaa 
tactacttt aaaaaaact tccaacaaa agactgagc 
ttggacgtg aacttgcgg ggcttcctc tgcagttgt 
aacaacagg gaaaaggac tgcattaag gaatcttac 
aatttacta aacgaatat gctgacaaa ttgtaccaa 
gagagcaac gaacctgaa gctgagcaa gacattgac 
gactcgttg gctaaagaa atatccgaa ctgaagcaa 
gacaagagt gaaaaacga ttccaggtt atagaatct 
ggtgccaaa aatttccta ttcattagg acgagtttg 
gagaatccg gttgagcta gcagaggcg attataaaa 
gatgttgat gggagtaag acccaaaga acgaaattt 
ttgctgaga ctgattcct gtggaaatt acgtgcaag 
gcgaatgtg agtgatatt gtcaacgca tttgtgccc 
ctcgcccag aaacacttt gtggagtcc ccgcaaacg 
ttttgtgtg atcttcaac caccggaat aataatgtt 
gtttcgcgg gatgaagtc ataaaatta attgcagcg 
aaagtgtcc gagctgagg cctgatcat aaagttgat 
cttaaagag gcaaaagtg gctattatt gttgaagtg 
ataaagggg tttgcgttt ttaagtgtt atacctgac 
tatcttaag cacaaaaag tataattta ttgtcactt 
tgtagtcaa gaacaagta gctgattgt aattaatgt 
tgcaataaa attttctat taatataaa aattttatt 
tccaaagtt ttgttctag aacagatac atggttgca 
ttgctacaa catgagtac acatagtgt aggtttgac 
tattaccat tgtttaaaa tttcaaagt tgcatcata 
aaattcaaa acattcgtc aatttcaag aagttaata 
aacattaaa gtaatatat ggtaaaagt ttaatatct 
ttatgctta atggtaaat aaatctttg tacaaata

    If you use iBeetle-Base, please cite: Dönitz J et al.: Expanded and updated data and a query pipeline for iBeetle-Base,
    Nucleic Acids Res. 2018 Jan 4;46(D1):D831-D835. doi: 10.1093/nar/gkx984.

    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY