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Gene information
General information
TC002852 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: 8144860..8145828
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
    		TC002852
    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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    iBeetle screen iB_00456

    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: 10.0% (includes death as pupa, adult)

    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): more than 80%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): more than 80%
    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%
    legs partially shape irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    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: 100.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)

    pupal molt delayed (still prepupa) - Number of animals showing the phenotype: 10 (11 dpi)
    Days post injection

    pupal molt not fulfilled (died as prepupa) - Number of animals showing the phenotype: 10 (19 dpi)
    Days post injection

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC002852 RA
      MDSQKTTFC NEISLMMFG FGDSHKPNP ETVRLVESI 
VLKQLRMIV QEALKYSDG KNLKGKELV FLMRHNKHK 
MRRFFQYLK NKQLKQQLQ SLNSGVIEI DRDPIPKNE 
LMKFIEYID ETGEFTDLS EVDEVKQSR MLRADRISQ 
ALDEKRYLE FCKARATSF CSRDITARN SEKLRQWID 
PKKEINFST NAMVVLSYL GYQTVAEIT DYALLVRME 
GKCGSDPLN HLYGNYYTS TMFNGDYRI GASGNNPDY 
SRVYSGQPP ISVNEIKEV MRRIHCPQA GKLNFGGKL 
PDTHYLFAL
    CDS sequence
    • >TC002852 RA
      atggattcc caaaaaaca accttttgt aacgaaatc 
tcgctgatg atgtttggg ttcggcgac agccacaag 
ccgaacccc gagacggtg cgtttggtg gagagcatc 
gtgttgaag cagctccgc atgatcgtc caggaagcg 
ctgaagtac tccgatggt aagaacctc aaaggcaag 
gagttggtg ttcctaatg cggcacaat aagcacaaa 
atgcgccgc tttttccaa tatttgaaa aacaagcaa 
ctcaaacag caactccag tcgttgaac tcgggggtc 
attgaaatt gacagagat cccataccg aagaacgag 
ctaatgaag tttatcgag tacatagac gagacggga 
gaattcacc gatttatcc gaagttgat gaagtcaaa 
cagtcgagg atgctgagg gctgatcgc atatctcaa 
gccctggac gagaagagg tatttggag ttttgcaaa 
gcgagagcc acctcgttt tgcagtaga gatatcacg 
gcccgaaac tccgaaaaa ttaagacag tggattgac 
ccgaaaaag gaaattaat ttcagcact aatgctatg 
gtggttttg tcgtatttg gggtaccag acagttgct 
gagataacc gattatgcg ttgcttgtg aggatggag 
ggcaaatgt gggagtgat ccactgaat catttatac 
ggcaattac tacacttct acgatgttt aatggggat 
tataggata ggggctagt ggtaacaat ccagactat 
agcagggtt tacagtgga cagcccccc attagtgtc 
aatgaaatt aaagaagtt atgagaaga attcactgt 
ccgcaagca gggaagctg aatttcggg ggaaaacta 
ccagacacg cattatcta tttgctctg taa
    mRNA sequence
    • >TC002852 RA
      aaaatattt gttgtcaat tgaggttag gtgtttgtt 
gacaaaaac tgatggatt cccaaaaaa caacctttt 
gtaacgaaa tctcgctga tgatgtttg ggttcggcg 
acagccaca agccgaacc ccgagacgg tgcgtttgg 
tggagagca tcgtgttga agcagctcc gcatgatcg 
tccaggaag cgctgaagt actccgatg gtaagaacc 
tcaaaggca aggagttgg tgttcctaa tgcggcaca 
ataagcaca aaatgcgcc gctttttcc aatatttga 
aaaacaagc aactcaaac agcaactcc agtcgttga 
actcggggg tcattgaaa ttgacagag atcccatac 
cgaagaacg agctaatga agtttatcg agtacatag 
acgagacgg gagaattca ccgatttat ccgaagttg 
atgaagtca aacagtcga ggatgctga gggctgatc 
gcatatctc aagccctgg acgagaaga ggtatttgg 
agttttgca aagcgagag ccacctcgt tttgcagta 
gagatatca cggcccgaa actccgaaa aattaagac 
agtggattg acccgaaaa aggaaatta atttcagca 
ctaatgcta tggtggttt tgtcgtatt tggggtacc 
agacagttg ctgagataa ccgattatg cgttgcttg 
tgaggatgg agggcaaat gtgggagtg atccactga 
atcatttat acggcaatt actacactt ctacgatgt 
ttaatgggg attatagga taggggcta gtggtaaca 
atccagact atagcaggg tttacagtg gacagcccc 
ccattagtg tcaatgaaa ttaaagaag ttatgagaa 
gaattcact gtccgcaag cagggaagc tgaatttcg 
ggggaaaac taccagaca cgcattatc tatttgctc 
tgtaatttg ttctaataa agtttaatt ttttaa
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY