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Gene information
General information
TC032266
Former TC numbers

If one gene is splitted or genes are merged, the original TC numbers are listed. One listed former TC number indicates the previous gene has be splitted, serveral former TC number represents a merged gene.

TC000189,TC000190 		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_007417.3: 14937469..14939249
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
    		TC032266
    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
    If you paste here the relevant section of the reference it is for us a great help to review your annotation. The quotation may be also be displayed to the users of iBeetle-Base.
    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_09709

    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: 1 (3 dpi)
    Days post injection

    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 embryo/egg with embryonic tissue, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): less than 30%

    embryo/egg with embryonic tissue, no cuticle, but muscle fluorescence - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    embryo/egg with muscle fluorescence/pattern, but not hatched (pot. late embryonic lethality) - Penetrance (percentage of animals/eggs on slide): less than 30%
    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): 50% - 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): less than 30%
    cuticle remnants with unidentifyable segments - Penetrance (percentage of animals/eggs on slide): less than 30%

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

    Comment: Constriction often occurs between thorax and abdomen segments.
    head appendages position irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    head not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    head shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%

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

    thorax not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    thorax shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    thorax not present - Penetrance (percentage of animals/eggs on slide): less than 30%
    abdomen not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    abdomen shape irregular - 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
    • >TC032266 RA
      MVIISKPLF YDKLELAIS CRELVHPDV GSCTKANLK 
LCSEDFLKG VFYFYLPIH LLQVILDYR KIKRKHFLK 
NLLQNVLRS ECFGVSFGA LIIATICLQ RHFTERFHY 
YSLLLLPGV VAGISLRWE HPRNRQLNT ILWASLVVE 
ALFKNAYYF KIFSYSRIK ETLIFMIFS SLLMHLFVN 
RDKKHIPFN LWFYKPSHP KKFKFTLWN YFIEISKYF 
GFGYGLNSV RCVLKDAKM VFKNPLQFV RKTIFNRRN 
VKFGIIIGA YVGLFRGVS VLLQKYTDF ERANAAIAG 
LVAGLSYYI QPNMTVLTT AIVTVFQIL CQQIATKPK 
FRKIPFAEL IFMFCNGIL FFNRIVAPV TCPKYYMNM 
LRITGNGEL EKIYVRLVN KYLP
    CDS sequence
    • >TC032266 RA
      atggttatt ataagcaaa ccgctattt tacgataaa 
cttgagctt gcaatatcg tgccgggag ttagttcat 
ccagatgtt ggcagttgt acgaaagcg aatctcaaa 
ctttgttcg gaggatttt ttaaaagga gtgttttat 
ttctattta cccattcat ttgttgcaa gttattctc 
gattataga aaaatcaag cgcaagcat tttctcaaa 
aatctgcta cagaacgtc ctaaggtct gaatgtttc 
ggtgtcagt tttggggca ctgataatt gccacaatc 
tgtctacaa cgtcatttt acagaacgg ttccattat 
tattctcta ttattactg ccaggtgta gtggcagga 
atttctcta cgatgggaa caccctaga aatcggcag 
ttaaatacg attctttgg gccagtttg gttgttgag 
gcattgttc aaaaatgct tactatttc aaaatattc 
agttattct cggataaaa gaaacgctt atttttatg 
atttttagt tctcttttg atgcattta tttgtaaac 
agggataag aaacacatt ccgttcaat ctttggttt 
tataaacca tcccacccg aaaaaattc aaattcacg 
ttgtggaat tattttatc gaaatttca aagtatttc 
ggattcggc tatggatta aatagtgtt aggtgcgtc 
ttgaaagac gccaaaatg gtgtttaaa aaccctctg 
caatttgtt agaaaaacg atctttaac agaagaaat 
gttaaattt ggaataatt attggtgct tacgttggt 
ttatttagg ggggtttca gttttactt cagaaatat 
acggacttt gaaagagca aatgctgca attgcagga 
cttgtagca ggattatct tactatatt cagccaaat 
atgaccgtt ttgaccacc gctattgta actgttttt 
caaatatta tgtcagcaa attgcaacc aaaccaaag 
tttcgaaaa attccgttt gctgaatta atttttatg 
ttttgtaat gggattctt ttcttcaat agaattgta 
gctccagtt acttgcccg aaatattac atgaatatg 
cttcgaatt actggaaat ggagaactg gaaaaaatc 
tacgttaga ctagtgaac aagtatttg ccataa
    mRNA sequence
    • >TC032266 RA
      taagtattt attcggtgt gttggaaac aatagtctg 
aataataaa aactaccaa cgacattat ttcgctgct 
tgagtccag taagcaaac atgaaagaa aaaagcgct 
tgaactgta ataattgtg actggtggc aaaatggtt 
attataagc aaaccgcta ttttacgat aaacttgag 
cttgcaata tcgtgccgg gagttagtt catccagat 
gttggcagt tgtacgaaa gcgaatctc aaactttgt 
tcggaggat tttttaaaa ggagtgttt tatttctat 
ttacccatt catttgttg caagttatt ctcgattat 
agaaaaatc aagcgcaag cattttctc aaaaatctg 
ctacagaac gtcctaagg tctgaatgt ttcggtgtc 
agttttggg gcactgata attgccaca atctgtcta 
caacgtcat tttacagaa cggttccat tattattct 
ctattatta ctgccaggt gtagtggca ggaatttct 
ctacgatgg gaacaccct agaaatcgg cagttaaat 
acgattctt tgggccagt ttggttgtt gaggcattg 
ttcaaaaat gcttactat ttcaaaata ttcagttat 
tctcggata aaagaaacg cttattttt atgattttt 
agttctctt ttgatgcat ttatttgta aacagggat 
aagaaacac attccgttc aatctttgg ttttataaa 
ccatcccac ccgaaaaaa ttcaaattc acgttgtgg 
aattatttt atcgaaatt tcaaagtat ttcggattc 
ggctatgga ttaaatagt gttaggtgc gtcttgaaa 
gacgccaaa atggtgttt aaaaaccct ctgcaattt 
gttagaaaa acgatcttt aacagaaga aatgttaaa 
tttggaata attattggt gcttacgtt ggtttattt 
aggggggtt tcagtttta cttcagaaa tatacggac 
tttgaaaga gcaaatgct gcaattgca ggacttgta 
gcaggatta tcttactat attcagcca aatatgacc 
gttttgacc accgctatt gtaactgtt tttcaaata 
ttatgtcag caaattgca accaaacca aagtttcga 
aaaattccg tttgctgaa ttaattttt atgttttgt 
aatgggatt cttttcttc aatagaatt gtagctcca 
gttacttgc ccgaaatat tacatgaat atgcttcga 
attactgga aatggagaa ctggaaaaa atctacgtt 
agactagtg aacaagtat ttgccataa aatcaaata 
gctaattag ttaattttt gtttacttt tttaacact 
aataaacct ctaacataa at

    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