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Gene information
General information
TC002951 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: 5859529..5860972
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
    		TC002951
    iBeetle screen iB_00476

    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: 90.0% (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: 3 (9 dpi)
    Days post injection

    adult lethality - Number of animals showing the phenotype: 5 (11 dpi)
    Days post injection

    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 20
    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: 90.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality early (L5/6) - Number of animals showing the phenotype: 9 (11 dpi)
    Days post injection

    larva lethality late (L7) - Number of animals showing the phenotype: 1 (19 dpi)
    Days post injection

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC002951 RA
      MAPTAEKKT KTVKGLPAV PESVLKHRK RRDAYRAKR 
LQTAIKRKS EQVKKRKEI FKRAERYVK EYRLKERDE 
IRLIRQAKT KGNFYVPGE AKLAFVIRI KGINKVAPK 
VRKVLQLFR LLQINNGVF VQLNKATIN MLRICEPYI 
TWGYPNLKS VRELIYKRG FAKVNGQRV PITSNQIIE 
NRLGKSEII CMEDLIHEI FTVGPKFKY ASNFLWPFK 
LNTPTGGWR KKTNHYVEG GDFGNREDK INELIRRMV
    CDS sequence
    • >TC002951 RA
      atggcgccg acggctgag aagaaaacg aagacagtt 
aaggggctg cctgccgtc cccgagtct gttttgaag 
caccggaag aggcgcgat gcgtaccgg gccaaacga 
ctccaaact gctatcaaa aggaagtcc gagcaggtg 
aaaaaacgc aaggaaatt tttaaacgc gccgaacga 
tacgtcaag gagtacagg ctcaaggaa cgtgatgaa 
attaggcta attcgtcaa gctaaaacc aagggtaac 
ttctatgtt cctggtgaa gccaagctg gctttcgta 
atccgtatc aagggtatt aacaaagtg gcccccaag 
gtgcgcaaa gtcctccaa ctgttccgc ttgctccaa 
atcaataat ggtgtcttt gtgcaactc aacaaggcc 
actatcaat atgttgaga atttgtgag ccttacatt 
acttggggt taccccaat ttgaagtca gtcagggaa 
ctgatctac aagagagga tttgccaaa gttaacggc 
cagcgcgtt ccaatcacc agcaatcaa atcattgaa 
aacagattg ggtaaatct gaaatcatc tgtatggaa 
gatttgatc catgaaatt tttactgtt ggacctaag 
tttaaatac gcctccaac ttcttgtgg cccttcaag 
cttaataca cccactggt ggatggcgc aagaagaca 
aaccactat gttgaggga ggtgatttt ggcaacagg 
gaagacaaa atcaatgaa ctcatcaga aggatggtt 
taa
    mRNA sequence
    • >TC002951 RA
      aagtagcct ccttttcca ctagatagt gatggcgcc 
gacggctga gaagaaaac gaagacagt taaggggct 
gcctgccgt ccccgagtc tgttttgaa gcaccggaa 
gaggcgcga tgcgtaccg ggccaaacg actccaaac 
tgctatcaa aaggaagtc cgagcaggt gaaaaaacg 
caaggaaat ttttaaacg cgccgaacg atacgtcaa 
ggagtacag gctcaagga acgtgatga aattaggct 
aattcgtca agctaaaac caagggtaa cttctatgt 
tcctggtga agccaagct ggctttcgt aatccgtat 
caagggtat taacaaagt ggcccccaa ggtgcgcaa 
agtcctcca actgttccg cttgctcca aatcaataa 
tggtgtctt tgtgcaact caacaaggc cactatcaa 
tatgttgag aatttgtga gccttacat tacttgggg 
ttaccccaa tttgaagtc agtcaggga actgatcta 
caagagagg atttgccaa agttaacgg ccagcgcgt 
tccaatcac cagcaatca aatcattga aaacagatt 
gggtaaatc tgaaatcat ctgtatgga agatttgat 
ccatgaaat ttttactgt tggacctaa gtttaaata 
cgcctccaa cttcttgtg gcccttcaa gcttaatac 
acccactgg tggatggcg caagaagac aaaccacta 
tgttgaggg aggtgattt tggcaacag ggaagacaa 
aatcaatga actcatcag aaggatggt ttaaaatgt 
taagacgag tttatataa taaattata taaaagtat
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY