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Gene information
General information
TC012015 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_007424.3: 8488610..8492409
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
    		TC012015
    iBeetle screen iB_05149

    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: 3 (3 dpi)
    Days post injection

    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 muscle fluorescence/pattern, but not hatched (pot. late embryonic lethality) - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    larval musculature complete not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    Comment: dead larva
    larval musculature partially not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    Comment: probably dying larva
    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
     thorax segment shape irregular - Penetrance (percentage of animals/eggs on slide): more than 80%

    thorax segment ventral size decreased - Penetrance (percentage of animals/eggs on slide): more than 80%

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

    larva lethality - Number of animals showing the phenotype: 10 (11 dpi)
    Days post injection

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC012015 RA
      MSYTVRQLS PEEAEVMDR AFGIKNSLM EYNPGKCLL 
PPFHDKIAQ QIIDAPVRE DDVWLISFP RTGSTWCQE 
MIWLIGNDL DFETARNTI QQIRAPLIE MSTVLIQYQ 
DTLGQELLG NSVDLVNNL PSPRYIKSH LPLPLLPTE 
LDKIKPKII YTCRNPKDM CVSYYHHCQ MFHQLDITF 
EEFCDQMIR GLTPMGALF PHYLSFWEK RHETNILFL 
KYEDMKKDL RGTLKKIAN FMEKSYTEE EYDKLCDFL 
SFQNMRDNR GCNLEVLLE SKYGKDYFK KTGKYFIRK 
GQVGDWKNH MSPELAKRF DDWIEENTR GTGLTFD
    CDS sequence
    • >TC012015 RA
      atgagttac acagtgagg caactaagt cccgaagaa 
gccgaagtt atggaccgg gccttcggg atcaagaac 
tccctcatg gagtacaat ccaggaaag tgcctcttg 
ccacccttc catgacaaa atcgcccaa caaatcatc 
gatgcacca gtgcgagaa gacgacgtt tggctcatt 
tccttccct cggacgggc tccacttgg tgccaggaa 
atgatctgg ctgattgga aatgacctc gactttgag 
acggcccga aacaccata cagcaaatc cgagctcct 
ttaatcgag atgtccaca gttctcatc caataccaa 
gacaccctc ggccaagaa ctcctcggc aactccgtc 
gacctcgtc aacaacctt ccgtcccca cgctacatc 
aaaagccac ctcccttta cctctactc cccacagaa 
cttgataaa atcaagccg aaaatcatc tacacttgt 
cgcaacccc aaggacatg tgcgtgtcg tattatcac 
cactgtcaa atgttccac cagcttgat atcacgttt 
gaggagttt tgcgaccag atgattaga gggttgacc 
ccaatgggt gccctcttt ccccattat ctctccttc 
tgggagaaa agacacgaa acgaatatt ctcttcttg 
aaatatgaa gatatgaag aaggatctt cgtgggact 
ttgaagaaa attgcaaat tttatggaa aagagttac 
actgaggaa gaatacgat aaattgtgc gactttttg 
agcttccaa aacatgcga gataaccga gggtgtaat 
ctcgaagtg ttgctagag tcgaaatat ggcaaggat 
tatttcaaa aagaccggg aaatatttt atcagaaag 
ggacaagtc ggggattgg aaaaaccac atgagtcca 
gaattggcc aaaaggttc gatgactgg attgaggaa 
aatacaaga gggaccgga ctcaccttt gattaa
    mRNA sequence
    • >TC012015 RA
      ctccagctg ctgcaacat catgagtta cacagtgag 
gcaactaag tcccgaaga agccgaagt tatggaccg 
ggccttcgg gatcaagaa ctccctcat ggagtacaa 
tccaggaaa gtgcctctt gccaccctt ccatgacaa 
aatcgccca acaaatcat cgatgcacc agtgcgaga 
agacgacgt ttggctcat ttccttccc tcggacggg 
ctccacttg gtgccagga aatgatctg gctgattgg 
aaatgacct cgactttga gacggcccg aaacaccat 
acagcaaat ccgagctcc tttaatcga gatgtccac 
agttctcat ccaatacca agacaccct cggccaaga 
actcctcgg caactccgt cgacctcgt caacaacct 
tccgtcccc acgctacat caaaagcca cctcccttt 
acctctact ccccacaga acttgataa aatcaagcc 
gaaaatcat ctacacttg tcgcaaccc caaggacat 
gtgcgtgtc gtattatca ccactgtca aatgttcca 
ccagcttga tatcacgtt tgaggagtt ttgcgacca 
gatgattag agggttgac cccaatggg tgccctctt 
tccccatta tctctcctt ctgggagaa aagacacga 
aacgaatat tctcttctt gaaatatga agatatgaa 
gaaggatct tcgtgggac tttgaagaa aattgcaaa 
ttttatgga aaagagtta cactgagga agaatacga 
taaattgtg cgacttttt gagcttcca aaacatgcg 
agataaccg agggtgtaa tctcgaagt gttgctaga 
gtcgaaata tggcaagga ttatttcaa aaagaccgg 
gaaatattt tatcagaaa gggacaagt cggggattg 
gaaaaacca catgagtcc agaattggc caaaaggtt 
cgatgactg gattgagga aaatacaag agggaccgg 
actcacctt tgattaact ttatctata gtacaaata 
ttttgttag ttgtgtata ttttttaat aaaattatt 
ttaaccta
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY