Status
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Gene information
General information
TC032448
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.

TC002799,TC002800 		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: 9678447..9749901
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
    		TC032448
    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_03452

    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: 40.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 4 (3 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): 30% - 50%

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

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - 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
     egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    cuticle remnants with some identifiable segments - Penetrance (percentage of animals/eggs on slide): less than 30%

    abdomen partially remaining - Penetrance (percentage of animals/eggs on slide): less than 30%
    procephalic appendages sometimes remaining - Penetrance (percentage of animals/eggs on slide): less than 30%
    head not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    thorax not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    abdomen not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    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: 0.0% (includes death as larva, prepupa, pupa, adult)

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC032448 RA
      MASSDLTLE QIYNYFKEN GNVVKNREL VHHFKHYLT 
DPASKEDAR VKFKKFVNT LAQTKTEGD EKFLILKTR 
YQNSLDYPR PSQLALSPH SDPRLALGI PLSPTPSDY 
SPAKSPAHR QPPPYRPPP PVTSPTNSL DNVSLSSIM 
SLQDVTPQA PPRRKQRSF DEDSTPRQI NNQEEEKQT 
PVSVKERLE KFNRMASME DELSPRQAK SAEKKKEKG 
SEEDDGASV TSLEPKKCM EWYVTASKG DIQELLKLA 
QDEPRLVNR KDPFTTVLH WGAKHGNPK IIQMFAGKY 
KVDVNGKTN GGYTPLHLA AQFGHRDIY DMLINEYHA 
DSKIRDYSG RLPAYYLQS KDQKIRKDN IRKIKGRKK 
PTDKDLGFL RIGSLNVRV KKTTEAFSN FLGVGSGSV 
TALETTSEK VHKGWGSAD NVNQENINP GPKINTVKK 
KSKRPYSSG MNSTPGTPR QMSRNFSSF HTNDSDSDS 
AAGFDSQWK H
    CDS sequence
    • >TC032448 RA
      atggcatcc agtgatttg acgttagaa caaatttac 
aattatttt aaagaaaat ggaaatgtg gttaaaaac 
cgtgaactc gttcatcac ttcaagcat tacttgacc 
gatcctgca tcaaaagaa gacgcccga gttaagttc 
aaaaaattc gtcaacacc ctcgcacaa accaaaacc 
gaaggcgac gaaaagttc cttatcctc aaaacccga 
taccaaaac tctctcgat taccctcga ccatcacaa 
ttggccctc tcaccccac agcgacccc cgactcgcc 
ctcggaatc ccactttcc ccaactcct tccgactac 
agtcctgcg aaatcacca gctcataga caacctcct 
ccttaccga cctccaccg ccggttact tccccaaca 
aactctctc gacaacgta tcgttaagt tcgataatg 
tcgctacaa gatgttaca ccacaagcg ccaccaaga 
cgcaaacaa aggtccttc gatgaggac tcaactccg 
cgacaaatt aacaaccaa gaggaggaa aaacaaacc 
ccggttagt gtcaaggag cggttggag aagtttaat 
cggatggcc tccatggag gacgagttg agtccgaga 
caggcgaag tcggcagag aagaagaag gaaaagggg 
tccgaagaa gacgacggc gcctcggtc acatcgtta 
gagccgaaa aagtgcatg gaatggtac gtaacggcg 
tcgaagggc gacatccaa gagctgctc aagctggcc 
caggacgag ccccgtctg gtcaacaga aaggatcca 
tttacgacg gttctgcat tggggcgcc aaacatggt 
aatccgaag atcatccaa atgttcgct ggcaagtac 
aaggtggat gtcaacggg aagacgaac gggggttac 
acaccacta cacctagca gcacagttc gggcacaga 
gatatttac gacatgctc ataaatgaa tatcatgct 
gattcgaaa attcgggac tatagcggc agattaccg 
gcttattac ttgcaaagt aaagatcaa aaaatccga 
aaagataac atccgcaag atcaaaggg cggaaaaaa 
cccacagat aaagatttg gggttttta cggatcggg 
tcgttgaat gtgcgagtt aaaaaaact acggaggct 
tttagcaat ttcttgggg gttggaagt ggttctgtc 
accgcactc gaaacgacc agtgagaag gtgcacaaa 
gggtggggc tctgccgat aatgtcaac caggaaaat 
ataaatccg gggcctaaa atcaacacg gtaaagaag 
aaatcgaaa aggccctat tcgtcagga atgaacagc 
actccgggg actccccgc caaatgtcg aggaatttt 
tccagtttt cacacaaat gactcggat tcggactca 
gccgccggg tttgactca caatggaag cactaa
    mRNA sequence
    • >TC032448 RA
      tgcttggaa ctgtatcga attatccaa gtgtagtgc 
ttggcgctt aattttgaa cgtggattg gtaaccgtg 
gcgcctata acggtaact tctagtaac tttcgccgt 
cagactgac gtttcgtgc gcggcatca tggcatcca 
gtgatttga cgttagaac aaatttaca attatttta 
aagaaaatg gaaatgtgg ttaaaaacc gtgaactcg 
ttcatcact tcaagcatt acttgaccg atcctgcat 
caaaagaag acgcccgag ttaagttca aaaaattcg 
tcaacaccc tcgcacaaa ccaaaaccg aaggcgacg 
aaaagttcc ttatcctca aaacccgat accaaaact 
ctctcgatt accctcgac catcacaat tggccctct 
caccccaca gcgaccccc gactcgccc tcggaatcc 
cactttccc caactcctt ccgactaca gtcctgcga 
aatcaccag ctcatagac aacctcctc cttaccgac 
ctccaccgc cggttactt ccccaacaa actctctcg 
acaacgtat cgttaagtt cgataatgt cgctacaag 
atgttacac cacaagcgc caccaagac gcaaacaaa 
ggtccttcg atgaggact caactccgc gacaaatta 
acaaccaag aggaggaaa aacaaaccc cggttagtg 
tcaaggagc ggttggaga agtttaatc ggatggcct 
ccatggagg acgagttga gtccgagac aggcgaagt 
cggcagaga agaagaagg aaaaggggt ccgaagaag 
acgacggcg cctcggtca catcgttag agccgaaaa 
agtgcatgg aatggtacg taacggcgt cgaagggcg 
acatccaag agctgctca agctggccc aggacgagc 
cccgtctgg tcaacagaa aggatccat ttacgacgg 
ttctgcatt ggggcgcca aacatggta atccgaaga 
tcatccaaa tgttcgctg gcaagtaca aggtggatg 
tcaacggga agacgaacg ggggttaca caccactac 
acctagcag cacagttcg ggcacagag atatttacg 
acatgctca taaatgaat atcatgctg attcgaaaa 
ttcgggact atagcggca gattaccgg cttattact 
tgcaaagta aagatcaaa aaatccgaa aagataaca 
tccgcaaga tcaaagggc ggaaaaaac ccacagata 
aagatttgg ggtttttac ggatcgggt cgttgaatg 
tgcgagtta aaaaaacta cggaggctt ttagcaatt 
tcttggggg ttggaagtg gttctgtca ccgcactcg 
aaacgacca gtgagaagg tgcacaaag ggtggggct 
ctgccgata atgtcaacc aggaaaata taaatccgg 
ggcctaaaa tcaacacgg taaagaaga aatcgaaaa 
ggccctatt cgtcaggaa tgaacagca ctccgggga 
ctccccgcc aaatgtcga ggaattttt ccagttttc 
acacaaatg actcggatt cggactcag ccgccgggt 
ttgactcac aatggaagc actaatacg aagcaacat 
cacggtgat tggacttaa atacgcttt attattttt 
atattatga tgtgtacct attttttaa taaattaat 
aaatttttg
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