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Gene information
General information
TC000466 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: 10923128..10924547
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
    		TC000466
    iBeetle screen iB_00100

    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: 30.0% (includes death as pupa, adult)

    pupa eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 3 (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): 50% - 80%

    condensed embryonic tissue, no cuticle, cns fluorescence - 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%
    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%
    larva cuticle fluorescence decreased - 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: 20.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 20.0% (includes death as larva, prepupa, pupa, adult)

    remarks: later than schedule, hours 100
    Adult morphology
    remarks: later than schedule, hours 100
    Fertility
    egg production: number no/almost no eggs -
    vitellogenic egg chamber late not present - Number of animals showing the phenotype: 4

    previtellogenic egg chamber potentially number decreased - Number of animals showing the phenotype: 4
    yolk amount decreased - Number of animals showing the phenotype: 4
    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC000466 RA
      MFLKVLIKN CSLIYYQRQ RDLSKLRRY GPVFIGGFA 
LFSSHKVKI DTSTFMASP ITPLSQLEQ APDDMKTRM 
ELMVLRVQA EACRALEEE DNCNKFLVD KWERHDGRG 
GGITCVLQE GKVFEKAGV NISVISGNL TPGIVAQMR 
ARGKKLKRD SPPFAVIGI SAVIHPKNP MVPTVHFNY 
RYFEIQDED QVVWWFGGG SDLTPYYLN KADAIHFHS 
ALKDACDKH DKSYYPKFK KWCDEYFTI KHRKERRGI 
GGIFFDDLD TPDRESCFK FATDCADAF IPSYIPLVQ 
RHKHAMYTE TERNWQLLR RGRYVEFNL VWDRGTKFG 
FNTPDARFE SILMSLPLL ARWEYMHQP APESPEGEL 
LEVLKNPRD WIEECSCTD HQKK
    CDS sequence
    • >TC000466 RA
      atgttcctc aaagtctta attaaaaac tgctcatta 
atttattac caacgtcaa cgcgactta tcaaagtta 
aggcgttat ggcccggtt tttatcgga ggcttcgcc 
ctattctct tcccacaaa gtaaaaata gacactagc 
acgttcatg gcatcgcct ataaccccc ttgtcgcaa 
ctggagcaa gcccccgat gacatgaag acccgtatg 
gagctgatg gtgctccgg gtccaggcg gaggcttgc 
cgagctctg gaggaggaa gacaattgt aacaagttc 
ctagtggat aagtgggag cgccacgac ggccgcggc 
ggtggtatc acttgtgtg ctccaagaa ggcaaagtt 
tttgagaaa gctggagtc aacatttcc gttatttcc 
ggaaatttg actccaggg attgtggca caaatgcgt 
gcaaggggg aagaaattg aaacgggat agtcccccg 
tttgccgtt atagggatc agtgctgtg attcacccc 
aaaaatccg atggtgcca actgttcat ttcaattac 
cggtatttt gaaatccag gatgaggat caggttgtg 
tggtggttt gggggcggg agtgacctc actccctac 
tatttgaat aaggctgat gctatacat ttccattcg 
gcgctcaag gatgcttgt gataaacat gacaaaagt 
tattatcct aagtttaag aagtggtgc gatgagtat 
ttcacgatt aaacatcgc aaggagagg aggggcatt 
ggggggata tttttcgat gatttggat actcccgat 
cgggagtcc tgtttcaaa tttgccacg gattgtgca 
gacgcgttc attccctcc tacattcct ttggtgcaa 
aggcataaa cacgccatg tacaccgag actgaaaga 
aactggcaa ttattgaga cggggacgt tacgttgag 
tttaacttg gtttgggat agaggtact aagttcgga 
tttaataca cctgatgct aggtttgaa agtatttta 
atgtcattg cctctcttg gcgagatgg gaatacatg 
caccaacca gctcccgaa tcacctgaa ggagagcta 
cttgaggtt ttaaaaaat cccagagac tggattgaa 
gagtgttct tgcacagac caccagaaa aaatga
    mRNA sequence
    • >TC000466 RA
      atgacatat ctaatctgc tatttttta ttttaacac 
tttccgact atcatttag tacaaaaac ttgccaatc 
agcatgttc ctcaaagtc ttaattaaa aactgctca 
ttaatttat taccaacgt caacgcgac ttatcaaag 
ttaaggcgt tatggcccg gtttttatc ggaggcttc 
gccctattc tcttcccac aaagtaaaa atagacact 
agcacgttc atggcatcg cctataacc cccttgtcg 
caactggag caagccccc gatgacatg aagacccgt 
atggagctg atggtgctc cgggtccag gcggaggct 
tgccgagct ctggaggag gaagacaat tgtaacaag 
ttcctagtg gataagtgg gagcgccac gacggccgc 
ggcggtggt atcacttgt gtgctccaa gaaggcaaa 
gtttttgag aaagctgga gtcaacatt tccgttatt 
tccggaaat ttgactcca gggattgtg gcacaaatg 
cgtgcaagg gggaagaaa ttgaaacgg gatagtccc 
ccgtttgcc gttataggg atcagtgct gtgattcac 
cccaaaaat ccgatggtg ccaactgtt catttcaat 
taccggtat tttgaaatc caggatgag gatcaggtt 
gtgtggtgg tttgggggc gggagtgac ctcactccc 
tactatttg aataaggct gatgctata catttccat 
tcggcgctc aaggatgct tgtgataaa catgacaaa 
agttattat cctaagttt aagaagtgg tgcgatgag 
tatttcacg attaaacat cgcaaggag aggaggggc 
attgggggg atatttttc gatgatttg gatactccc 
gatcgggag tcctgtttc aaatttgcc acggattgt 
gcagacgcg ttcattccc tcctacatt cctttggtg 
caaaggcat aaacacgcc atgtacacc gagactgaa 
agaaactgg caattattg agacgggga cgttacgtt 
gagtttaac ttggtttgg gatagaggt actaagttc 
ggatttaat acacctgat gctaggttt gaaagtatt 
ttaatgtca ttgcctctc ttggcgaga tgggaatac 
atgcaccaa ccagctccc gaatcacct gaaggagag 
ctacttgag gttttaaaa aatcccaga gactggatt 
gaagagtgt tcttgcaca gaccaccag aaaaaatga 
ttgtttttt taaataatt ttgataaat aaat
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY