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Gene information
General information
TC004532 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: 3054103..3055307
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
    		TC004532
    iBeetle screen iB_00714

    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: 20.0% (includes death as pupa, adult)

    adult lethality - Number of animals showing the phenotype: 2 (11 dpi)
    Days post injection

    adult nutritional state bad (cachectic) - Number of animals showing the phenotype: 4 (11 dpi)
    Days post injection

    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: 10.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 75.0% (includes death as larva, prepupa, pupa, adult)

    pupal molt not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection

    eclosion not fulfilled (not completely eclosed) - Number of animals showing the phenotype: 3 (16 dpi)
    Days post injection

    adult lethality immediately after eclosion - Number of animals showing the phenotype: 2 (19 dpi)
    Days post injection

    pupal molt not fulfilled (died as pupa) - Number of animals showing the phenotype: 3 (19 dpi)
    Days post injection

    adult lethality - Number of animals showing the phenotype: 1 (22 dpi)
    Days post injection

    Pupal morphology
    wings shortened - Number of animals showing the phenotype: 9

    wings not closed - Number of animals showing the phenotype: 9
    Fertility
    egg production: number no/almost no eggs -
    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC004532 RA
      MLFLRKLVS PSPCVCVNF VRHAKRIVD PGIRRRKEL 
LQLDESEEI DDFDDFETD FMEVHKSHK QHLREAEVE 
REKMKYHIV KQRYFKEKF PNFLTWHDK EQIRYLYRT 
DPNEWTIER LSESFPALP EVIKKIIKS NWDKPSQNK 
ITNHDTAVQ NNWKLFKNG KMGDLPADL AQHLNKFTN 
RALNLNQFK LPDFKTNVV TVGKKIGTE FSDIVTSYG 
GSHQNEDEK LEIEAKTLP DKKSKSRKP VTLNQLQVK 
LVKRAELGK QIDENEKCI VSSVKTEEE KIEVKHDLV 
DLSHNKFQT GVQLHKKEE KDLSHLNYP EVITIPKSA 
LKKGCIYKL NDCYYDDDG EFLYRVPGM SQ
    CDS sequence
    • >TC004532 RA
      atgttattt ctgagaaaa ctagtttca ccttctccc 
tgcgtttgt gttaatttt gtccgacat gcgaagaga 
atcgttgat cccggaata cgaagacgc aaggaatta 
ttacaactc gacgaatcc gaagaaatt gacgatttt 
gacgacttt gaaaccgat tttatggaa gtacacaag 
tcccataaa caacacctt cgagaagca gaggttgag 
agggaaaaa atgaaatat cacattgta aagcaacgg 
tattttaaa gaaaagttt ccgaatttt ttaacttgg 
catgataag gaacaaatt cggtatttg taccgtact 
gatccgaat gagtggacg attgaacgg ttatctgag 
agttttcca gcattgccg gaagtcatc aagaaaatt 
atcaaatca aattgggac aaaccaagt cagaataaa 
attacaaat cacgacact gcggtccaa aataactgg 
aagttgttt aaaaacggg aaaatgggg gatttacct 
gcggatttg gcccaacat ttgaataag tttacaaat 
cgcgcactc aatttgaat cagtttaaa ttaccagac 
tttaaaacc aatgtagtg actgttggg aaaaaaatt 
ggaacggag ttttctgat attgtgact agttacggt 
ggtagccac caaaatgag gatgaaaaa ctagaaata 
gaagcaaaa actttgccc gataaaaaa tcaaaaagt 
cgaaaacct gtcacactt aatcagctt caagtaaaa 
ctggtcaaa agggcagaa ttaggtaag caaatcgat 
gagaacgaa aaatgcata gtaagcagt gtgaagacg 
gaggaagag aaaattgaa gtaaaacac gacttggtt 
gatttgtcc cacaacaaa tttcaaact ggggtccaa 
cttcacaaa aaagaggaa aaggatttg tctcatttg 
aattacccg gaagttatc accataccc aaatctgcg 
ctaaagaaa ggctgtatt tacaaactc aatgattgt 
tactacgac gatgacggc gaatttttg tatcgagtc 
ccaggaatg tcgcaataa
    mRNA sequence
    • >TC004532 RA
      aaattttta cgtccataa acctcactt ttttaatta 
ataatttaa aacgatgtt atttctgag aaaactagt 
ttcaccttc tccctgcgt ttgtgttaa ttttgtccg 
acatgcgaa gagaatcgt tgatcccgg aatacgaag 
acgcaagga attattaca actcgacga atccgaaga 
aattgacga ttttgacga ctttgaaac cgattttat 
ggaagtaca caagtccca taaacaaca ccttcgaga 
agcagaggt tgagaggga aaaaatgaa atatcacat 
tgtaaagca acggtattt taaagaaaa gtttccgaa 
ttttttaac ttggcatga taaggaaca aattcggta 
tttgtaccg tactgatcc gaatgagtg gacgattga 
acggttatc tgagagttt tccagcatt gccggaagt 
catcaagaa aattatcaa atcaaattg ggacaaacc 
aagtcagaa taaaattac aaatcacga cactgcggt 
ccaaaataa ctggaagtt gtttaaaaa cgggaaaat 
gggggattt acctgcgga tttggccca acatttgaa 
taagtttac aaatcgcgc actcaattt gaatcagtt 
taaattacc agactttaa aaccaatgt agtgactgt 
tgggaaaaa aattggaac ggagttttc tgatattgt 
gactagtta cggtggtag ccaccaaaa tgaggatga 
aaaactaga aatagaagc aaaaacttt gcccgataa 
aaaatcaaa aagtcgaaa acctgtcac acttaatca 
gcttcaagt aaaactggt caaaagggc agaattagg 
taagcaaat cgatgagaa cgaaaaatg catagtaag 
cagtgtgaa gacggagga agagaaaat tgaagtaaa 
acacgactt ggttgattt gtcccacaa caaatttca 
aactggggt ccaacttca caaaaaaga ggaaaagga 
tttgtctca tttgaatta cccggaagt tatcaccat 
acccaaatc tgcgctaaa gaaaggctg tatttacaa 
actcaatga ttgttacta cgacgatga cggcgaatt 
tttgtatcg agtcccagg aatgtcgca ataaataaa 
agaaataaa attgtgtat ttttt
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