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Gene information
General information
TC004658 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_007416.3: 1028687..1031504
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
    		TC004658
    iBeetle screen iB_09962

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

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

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    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): 50% - 80%
    cuticle remnants with unidentifyable segments - Penetrance (percentage of animals/eggs on slide): less than 30%

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

    head not present - Penetrance (percentage of animals/eggs on slide): less than 30%
    thorax shape irregular - Penetrance (percentage of animals/eggs on slide): less than 30%
    thorax not present - Penetrance (percentage of animals/eggs on slide): less than 30%
    abdomen shape irregular - Penetrance (percentage of animals/eggs on slide): less than 30%
    abdomen not present - Penetrance (percentage of animals/eggs on slide): less than 30%
    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
    • >TC004658 RA
      MPKKFSTEN TKAVAARER KKAAKDAEV 
AKKQQQLEE AYWQDDDKL IKKKQQRKE 
EQERKREQQ LAKKQEAKQ LLEQEMSSI 
KKAGKPAPP PKVTRSQIS DTIAKKEPE 
KKEKVVTHL DEPLQENIN RLKIEGEEA 
RTVDEAIGI LSSKPDDID KHPERRMKS 
AFTAYETRR LEELKAENP TLRLSQLKQ 
MIFKEWQKS PENPLNKQ
    CDS sequence
    • >TC004658 RA
      atgccaaaa aaattctcc accgaaaat accaaagct 
gtggcagca cgcgaacgt aaaaaagct gcaaaagac 
gccgaagtg gccaaaaaa cagcaacaa ttagaagaa 
gcctactgg caagatgac gacaaatta atcaaaaag 
aagcaacag agaaaggag gagcaagaa cgtaaaagg 
gagcaacaa ttagccaaa aaacaggaa gccaagcag 
ctcctggaa caggaaatg agctctatt aaaaaagcc 
ggaaagcct gcgccgcca cccaaagtg acacgatcc 
caaatttcg gacaccata gccaaaaaa gagccagaa 
aagaaggaa aaagttgta acgcatctg gacgaaccc 
ttgcaggaa aacatcaac aggctcaag attgagggg 
gaggaggcc aggactgtg gacgaggcc attggaata 
ttgagctcg aaaccggat gatatcgac aagcacccc 
gagcgtcgg atgaagtct gcatttaca gcgtacgag 
acgagacgt ctggaggaa ctgaaggct gaaaacccc 
acacttcgc ctgtcccag ttgaagcaa atgatattc 
aaagagtgg caaaaatcg ccggagaat cctctgaat 
aaacaataa
    mRNA sequence
    • >TC004658 RA
      ggttgaggt tacatattt atttacaaa agtattact 
ttacattta cattgaatg acaaaaaaa tgccaaaaa 
aattctcca ccgaaaata ccaaagctg tggcagcac 
gcgaacgta aaaaagctg caaaagacg ccgaagtgg 
ccaaaaaac agcaacaat tagaagaag cctactggc 
aagatgacg acaaattaa tcaaaaaga agcaacaga 
gaaaggagg agcaagaac gtaaaaggg agcaacaat 
tagccaaaa aacaggaag ccaagcagc tcctggaac 
aggaaatga gctctatta aaaaagccg gaaagcctg 
cgccgccac ccaaagtga cacgatccc aaatttcgg 
acaccatag ccaaaaaag agccagaaa agaaggaaa 
aagttgtaa cgcatctgg acgaaccct tgcaggaaa 
acatcaaca ggctcaaga ttgaggggg aggaggcca 
ggactgtgg acgaggcca ttggaatat tgagctcga 
aaccggatg atatcgaca agcaccccg agcgtcgga 
tgaagtctg catttacag cgtacgaga cgagacgtc 
tggaggaac tgaaggctg aaaacccca cacttcgcc 
tgtcccagt tgaagcaaa tgatattca aagagtggc 
aaaaatcgc cggagaatc ctctgaata aacaataat 
gtcgcaacc aagtgatca tcatcgtga atatgccga 
aataatttc ttacgcact aacctactt atacttgac 
gctttattg ttttaccgg ttttattag acgaattgt 
tatgaagtc agtgtattt tgttggtac cgttaatag 
ataatttat gacacacaa atatttcat ttcattgca 
taaccctat aaatcatgt cgcgtttcg tgtgggtaa 
ggctggagg agggacgtc gggaatgtt tgggtttct 
cttcaggtt tgcgatctc gtgtttagg atcatcggc 
tttttggta aatttcttc gggtagtgt ttttggctt 
ttcgggctg agtttcctg cgcgtagta gcccggagg 
ggcgtaggc gtatggtac gaaccgctc cacaggtag 
tggttgtag cgcactttg taactcctc attttctgg 
tttgcctac tttcggtgg tgtcgcaac cttaaaata 
acaactcgt gctgttctc ctggcagag tctcactta 
cgaaggtgg cgatcggcg attgataga aaaagggcg 
gtagtgcta ttccgaggg atgttgtcc tcaacaccg 
gctcgtaga cggggaggg catgactag gggataaat 
tcttcttcg tgcttatca aaacgttgt atatctttg 
gggttctcg gatcagcac gtaccagga ataacagtg 
ctgacaatg aaaacaatc tgtacatag tatctgcaa 
aaagacatt gaattaaag gtcaccgcc ttcatcaag 
cagatctcc gaatttttt aggtaaacc tgtttgagt 
aatgatgca gtgagacta atctctagt tacggttgg 
ctacgctct tcagtaacg tggattcaa tgattttcg 
caggaactg agtaatgct gaatttaat ttttgccaa 
accgcacgt cccgattat aattgcaaa tcggagact 
gtaattgat ccagttttt cggctgcgt gttaattac 
accaagggt gattaattg ttaatttac acaggccac 
tgaatggac gatttatga aaaggtcgc cggtggact 
tagcgcact tggaaaaat tgaatgaac actagcaca 
tgttattgt ttaccttgt ataaattgt ttcacgtca 
caactgcaa ataattgcc ttgatagaa cagagaggt 
cttatatag ttcttgagg ttgttcttt gtgccgctc 
aactattcg tgttcactt gtgaagtac atttttcag 
ataaatccc ttcactggg agccttgca tacgaccaa 
ggtatttgc aattaatta cgttgttgt gatcactcc 
gagtaaatt ttcccacgg atgaatgaa aattttttt 
gatggacct gacttaggc gtgggcctg atgaaatat 
tccaaggaa aaaggcatt agccaactg gaaggagtg 
tcggaaatc atccgagcc aggtgcaca ttgtctcag 
taccaagcc tcttaccaa ttca
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY