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Gene information
General information
TC000350 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: 12716193..12717493
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
    		TC000350
    iBeetle screen iB_06818

    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: 70.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 2 (3 dpi)
    Days post injection

    adult lethality - Number of animals showing the phenotype: 5 (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 potentially present, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    larval musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    remarks: muscle pattern potentially obscured by segmentation defect,
    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): 30% - 50%
    cuticle remnants with unidentifyable segments - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    head capsule mostly not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    antenna not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    labrum 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 cuticle structure irregular -

    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
    • >TC000350 RA
      MASFSVINL QGYEAIIHE KLFANIKQL TYVYPPTQD 
FHEVFREDM FKKNNKAAF QQVTNYLFN VLSPELTRE 
KLKSWPPYD TKGEREFRQ EVVAFVDYL NENYKAANI 
PKLMPSLFV SPGGIKVAK FMFKLSTFV LHEHLKKNM 
KEGKLLSPI RPCKNPAVN AIIIENLNR TTSVIKKRT 
SETATNFHK FSAEAFAEA QRIVEQVER IKRQEMAEV 
TKALDEEKK KNLESIDLE KLQGKMAQM ENIKSMCER 
ANELLDVIM NDNQLELDK ENTPDLDLL QLFNDLNEF 
LEKERFQIP TLLQEDMKD MVQRLSELT ENYKQLIIN 
MEHDKKELE KLAENSVI
    CDS sequence
    • >TC000350 RA
      atggcgtca ttctctgtg ataaatttg caaggttac 
gaagccata atccacgag aaacttttc gcaaacatt 
aagcaatta acttatgtt tatccacca acccaagac 
ttccatgag gttttcagg gaagatatg tttaaaaaa 
aataacaag gcagccttc caacaagtc accaactac 
cttttcaat gtcctaagt cctgagttg accagagaa 
aagctcaaa tcctggccc ccatacgac accaaagga 
gagcgagag tttagacaa gaggtggtg gcatttgtt 
gattacttg aatgaaaat tacaaagcg gcgaacatt 
cctaagcta atgccatct ctttttgtt tcacctggt 
ggcataaag gtggccaaa ttcatgttt aaattgtcg 
acgttcgtc ttgcatgag catttaaaa aagaacatg 
aaagagggc aaattattg tctcccatc agaccttgt 
aaaaatcca gccgttaat gcaataatt atagaaaac 
ttgaataga acaacgtca gttataaaa aaaagaacg 
tcagaaact gccaccaac ttccacaaa ttctcggca 
gaggcattt gcagaagcc cagcgaatt gtggagcaa 
gtggagagg attaaaaga caggaaatg gccgaagtc 
accaaggcg ttagacgaa gagaaaaag aaaaatctc 
gaatcaatt gatttggaa aagttgcaa ggaaaaatg 
gcacaaatg gaaaatatt aaaagcatg tgtgagaga 
gcaaacgag ctcctggat gtcataatg aatgataat 
caactggaa cttgataaa gaaaatact cccgacttg 
gatttgctg cagttgttc aatgattta aatgagttc 
cttgaaaag gaacgtttc caaatacca actttgtta 
caggaagat atgaaagat atggttcaa cgcttgtca 
gagctcaca gaaaactac aaacaactt atcattaat 
atggagcac gacaagaag gaactcgaa aaattggca 
gaaaattct gttatctaa
    mRNA sequence
    • >TC000350 RA
      attaaaata aaaaaacag ttaaaaaac gtctgtttt 
taataaaaa aatcgtccg tctggtgat ttatttttc 
tgacgttgg tcgtggtca attaaagtt actggtctt 
tagtttaaa aaagtcgtg tttttgaag cttatttgt 
tctttcaca gaatggcgt cattctctg tgataaatt 
tgcaaggtt acgaagcca taatccacg agaaacttt 
tcgcaaaca ttaagcaat taacttatg tttatccac 
caacccaag acttccatg aggttttca gggaagata 
tgtttaaaa aaaataaca aggcagcct tccaacaag 
tcaccaact accttttca atgtcctaa gtcctgagt 
tgaccagag aaaagctca aatcctggc ccccatacg 
acaccaaag gagagcgag agtttagac aagaggtgg 
tggcatttg ttgattact tgaatgaaa attacaaag 
cggcgaaca ttcctaagc taatgccat ctctttttg 
tttcacctg gtggcataa aggtggcca aattcatgt 
ttaaattgt cgacgttcg tcttgcatg agcatttaa 
aaaagaaca tgaaagagg gcaaattat tgtctccca 
tcagacctt gtaaaaatc cagccgtta atgcaataa 
ttatagaaa acttgaata gaacaacgt cagttataa 
aaaaaagaa cgtcagaaa ctgccacca acttccaca 
aattctcgg cagaggcat ttgcagaag cccagcgaa 
ttgtggagc aagtggaga ggattaaaa gacaggaaa 
tggccgaag tcaccaagg cgttagacg aagagaaaa 
agaaaaatc tcgaatcaa ttgatttgg aaaagttgc 
aaggaaaaa tggcacaaa tggaaaata ttaaaagca 
tgtgtgaga gagcaaacg agctcctgg atgtcataa 
tgaatgata atcaactgg aacttgata aagaaaata 
ctcccgact tggatttgc tgcagttgt tcaatgatt 
taaatgagt tccttgaaa aggaacgtt tccaaatac 
caactttgt tacaggaag atatgaaag atatggttc 
aacgcttgt cagagctca cagaaaact acaaacaac 
ttatcatta atatggagc acgacaaga aggaactcg 
aaaaattgg cagaaaatt ctgttatct aataaagat 
tgaaattac tttacattt tttgtacca taataataa 
aaataaatt ttttgtatt acc
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY