Status
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Gene information
General information
TC000316 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: 13085514..13093033
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
    		TC000316
    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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    Publish name
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    * : mandatory fields
    For help on how to submit larger datasets or non experimental data please contact us.
    iBeetle screen iB_00069

    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: 0.0% (includes death as pupa, adult)

    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 - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 50
     thorax partially constricted - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): less than 30%
    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: 10.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 60.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality early (L5/6) - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection

    adult lethality immediately after eclosion - Number of animals showing the phenotype: 5 (19 dpi)
    Days post injection

    Adult morphology
    motility irregular - (22 dpi)
    Days post injection
    Comment: Legs are correctly formed, but can't be streched out. Joints between femur and tibia are not moveable.  

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC000316 RA
      MWRLLGFGV VLAAVAAPN NQVPKTGFS CEGRTTGYY 
ADVESGCQV YHMCDGLGR QFSYTCPNA TLFQQRMLI 
CDHWYMVNC SKSVDDYTA NLRIGHKEM PFVDDNEAN 
PYHRTPRPD LLSHPSQSE YDIIYRTGR AQLGTNLNL 
VGAESDPKN STTSTTDEP AYSLPSHWS TEYNKQVTT 
TKPKPAKKG KPTTVRVNY QSNYKATTP VFPQSVEVT 
EAPDLELVP PSGSTESSV NFESRFKAT TPVFPLSVD 
VTEAPNLGL LPPLPYQLN DTKPNENLV VSTVNFESK 
FKATTPVYP KSVELTSEE PSEVGVLPP KDTNSTETD 
VVPPLSIDF EPPVVDSNF KPGDHSEEI PPELPSKFY 
QPPKFEPDY TELAKLKGG FKALSSGEW EDLRKKFLI 
PDYEFPLET VSRPSYDSV LSSFQVGPV PSEKE
    CDS sequence
    • >TC000316 RA
      atgtggcgg cttttagga tttggagtc gttctggcc 
gccgtcgcg gcgcccaac aaccaggtt cctaagact 
ggcttttcc tgcgagggc cgcaccacg gggtactac 
gccgacgtg gaaagcggc tgccaagtc taccacatg 
tgtgacggt ttaggccgc cagttcagc tacacttgt 
cccaatgca actctcttc caacagaga atgttgatt 
tgcgaccac tggtacatg gtcaactgt agcaaatct 
gtggatgat tacacggcc aatttgcgc atcgggcac 
aaggaaatg cccttcgtt gacgacaac gaagccaat 
ccttaccac aggaccccc aggcctgac ctcctctcg 
caccccagt cagtccgaa tacgacatt atctaccgc 
accggaagg gcgcagctt ggcaccaat ctgaatcta 
gtcggcgcc gaatcagac cccaaaaac agtaccacc 
agtaccaca gacgaaccc gcctattct ctacccagc 
cactggtca acggaatat aacaaacaa gtgaccaca 
actaaacca aaaccggcc aaaaagggg aaacccacc 
acagtcagg gtgaactac cagtctaat tataaagcc 
acgactcct gtgttcccc caaagtgtg gaggtgacc 
gaagccccc gatttggaa ctagtgccc ccttcgggg 
agtaccgaa agctcggtt aatttcgag tcgaggttc 
aaggctacg actccggtg tttccccta agtgtggac 
gtgacggaa gccccaaat ttgggactt ctgccacct 
ctaccctac caacttaac gacacgaaa ccaaacgaa 
aatttagtc gttagtacg gtcaatttc gagtcaaaa 
ttcaaggcc acgacccca gtctacccc aaaagtgtc 
gaactgacg agtgaagaa ccgtccgaa gttggggtt 
ttgcccccc aaagacacc aattcgact gaaactgat 
gttgtgccc cctttaagt atagatttt gagccgcct 
gttgtcgac tctaatttc aagccaggt gatcacagt 
gaggaaata ccaccagaa ttaccgtca aagttctac 
caacccccg aaatttgaa cctgactat accgaatta 
gcgaagcta aagggtggg tttaaggcg cttagcagc 
ggcgaatgg gaggatttg aggaagaag ttcctgatt 
cccgattac gagtttccc ttagagacc gtttcaagg 
cctagttat gacagtgtt ttgagttcg tttcaagtc 
ggaccagtg ccatccgag aaagagtga
    mRNA sequence
    • >TC000316 RA
      gccgagttg tagatttgt agtgggacg ggttgtatc 
gtgaagtgt gaagaacgg tacctgaaa tgtggcggc 
ttttaggat ttggagtcg ttctggccg ccgtcgcgg 
cgcccaaca accaggttc ctaagactg gcttttcct 
gcgagggcc gcaccacgg ggtactacg ccgacgtgg 
aaagcggct gccaagtct accacatgt gtgacggtt 
taggccgcc agttcagct acacttgtc ccaatgcaa 
ctctcttcc aacagagaa tgttgattt gcgaccact 
ggtacatgg tcaactgta gcaaatctg tggatgatt 
acacggcca atttgcgca tcgggcaca aggaaatgc 
ccttcgttg acgacaacg aagccaatc cttaccaca 
ggaccccca ggcctgacc tcctctcgc accccagtc 
agtccgaat acgacatta tctaccgca ccggaaggg 
cgcagcttg gcaccaatc tgaatctag tcggcgccg 
aatcagacc ccaaaaaca gtaccacca gtaccacag 
acgaacccg cctattctc tacccagcc actggtcaa 
cggaatata acaaacaag tgaccacaa ctaaaccaa 
aaccggcca aaaagggga aacccacca cagtcaggg 
tgaactacc agtctaatt ataaagcca cgactcctg 
tgttccccc aaagtgtgg aggtgaccg aagcccccg 
atttggaac tagtgcccc cttcgggga gtaccgaaa 
gctcggtta atttcgagt cgaggttca aggctacga 
ctccggtgt ttcccctaa gtgtggacg tgacggaag 
ccccaaatt tgggacttc tgccacctc taccctacc 
aacttaacg acacgaaac caaacgaaa atttagtcg 
ttagtacgg tcaatttcg agtcaaaat tcaaggcca 
cgaccccag tctacccca aaagtgtcg aactgacga 
gtgaagaac cgtccgaag ttggggttt tgcccccca 
aagacacca attcgactg aaactgatg ttgtgcccc 
ctttaagta tagattttg agccgcctg ttgtcgact 
ctaatttca agccaggtg atcacagtg aggaaatac 
caccagaat taccgtcaa agttctacc aacccccga 
aatttgaac ctgactata ccgaattag cgaagctaa 
agggtgggt ttaaggcgc ttagcagcg gcgaatggg 
aggatttga ggaagaagt tcctgattc ccgattacg 
agtttccct tagagaccg tttcaaggc ctagttatg 
acagtgttt tgagttcgt ttcaagtcg gaccagtgc 
catccgaga aagagtgaa ggggaaaaa gagtttttt 
cctttaaag tctgatacc aaagttggg taatagcat 
gagatactc atttttcaa attatgatt gtaaataat 
agatccgtt gtgtttaat aacagttgg aaaaaccag 
tcaattata atgccacaa ccattgtta caagcgtct 
gatacgtgt aaaatgcgc acgatgaaa tagctttaa 
ttaaatcgg gttatataa ctgtgtcgc cattttttt 
aaggactca cgtttaatt tatactttt gtaaaacgt 
aattaaata aagaatgtt aagagga
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY