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Gene information
General information
TC030232 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_007425.3: 2137622..2139051
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
    		TC030232
    iBeetle screen iB_08338

    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 (9 dpi)
    Days post injection

    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): less than 30%

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

    larval musculature fluorescence decreased - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 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%
    head not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    head 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%
    thorax 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
    • >TC030232 RA
      MQALTLTSI FAPQHVVSK ILGVLPFTI QDGRPVFSK 
LGLCHSFLC IILYVFVAQ SYYQTIFDD TTSGFITLK 
ILKFITIAR YVGYTAVMI NTFVAVGLT YKKFVALLS 
KLYGIDRGL GKLGLETFV VASGYRIKK KMFLLVILE 
YLIGNIVSI FSSQFNDIS GAFVLLYPR MIGCNLSVT 
YFGTTLLLE ERFKLVNKF LLGECFKSR NCTKYPTDP 
NFCDKIKLS VNIHKELTR ICRAINSIF SVFFLLWVT 
VNFLCLVSD LYIVAYLIS SGQFFNYWQ GELKAFKNV 
CIYSIEMFY IARRSSRLC SQANRTKTI LVGLKVDLS 
KEEERNTII TSSLELRQF TLQITACRL FTIDNALLF 
SICGAAFSY LFIMIQMDK DLNKSLRNV TNSIL
    CDS sequence
    • >TC030232 RA
      atgcaagcg ctcactctg acgtcgatt tttgcaccc 
caacatgtc gtttcaaaa attttgggt gtactaccc 
ttcacaata caggacggt cgacctgtg ttttccaaa 
cttggtctt tgtcacagt tttctgtgc atcattttg 
tacgttttt gtggcccaa agctactac cagacaata 
tttgatgat acgacctca ggttttatt acattaaaa 
attttgaaa tttattaca atcgcccgc tatgttggt 
tacacggcc gtgatgatt aacacattc gttgctgtt 
ggtctgact tacaaaaaa tttgtcgct ttgttgtcg 
aaattgtat ggaattgat cgaggattg gggaagttg 
ggtctagag acatttgtt gtagcaagt ggttacagg 
attaagaaa aaaatgttc ttgttggtg attttggag 
tatttaatt ggaaatata gtgtcgatt ttttcatcg 
caattcaac gacatttcc ggtgctttt gttttactt 
taccctcga atgattggt tgtaacttg agtgtgacc 
tattttggt accactttg ttactggaa gagagattc 
aaattggtc aataagttc ctacttggg gagtgtttc 
aagtcacga aattgtacc aaatatcca accgatcca 
aatttttgc gacaaaatc aaattatcg gtcaatatt 
cacaaagaa ctgaccaga atttgtaga gctatcaac 
tcgattttt tcagttttt tttctgctt tgggtcact 
gtgaatttc ttgtgcttg gtttcagat ctgtacatt 
gttgcttat ttaatctca agcggccag ttttttaat 
tattggcaa ggcgagttg aaagctttt aagaacgtt 
tgtatttat tcgattgag atgttttat attgcgaga 
agatcgtca cggctttgt tctcaagca aacagaact 
aaaactatt ttggttggg ttgaaagtg gatttaagc 
aaagaggag gaacgaaat actattatt acttcgagt 
cttgaactt cgacagttt actcttcaa ataacggct 
tgtagactt tttacgatt gataatgcg ttacttttt 
tcgatctgt ggagctgcc ttttcatac ctcttcata 
atgatacaa atggacaaa gacttgaat aagtctctc 
cgaaatgta acaaattct attctatag
    mRNA sequence
    • >TC030232 RA
      gacaaatac aatcaataa cccattatt ataattttc 
aaatggttg aatttggtt tttaatgcc agttcaact 
gaagcaatg caagcgctc actctgacg tcgattttt 
gcaccccaa catgtcgtt tcaaaaatt ttgggtgta 
ctacccttc acaatacag gacggtcga cctgtgttt 
tccaaactt ggtctttgt cacagtttt ctgtgcatc 
attttgtac gtttttgtg gcccaaagc tactaccag 
acaatattt gatgatacg acctcaggt tttattaca 
ttaaaaatt ttgaaattt attacaatc gcccgctat 
gttggttac acggccgtg atgattaac acattcgtt 
gctgttggt ctgacttac aaaaaattt gtcgctttg 
ttgtcgaaa ttgtatgga attgatcga ggattgggg 
aagttgggt ctagagaca tttgttgta gcaagtggt 
tacaggatt aagaaaaaa atgttcttg ttggtgatt 
ttggagtat ttaattgga aatatagtg tcgattttt 
tcatcgcaa ttcaacgac atttccggt gcttttgtt 
ttactttac cctcgaatg attggttgt aacttgagt 
gtgacctat tttggtacc actttgtta ctggaagag 
agattcaaa ttggtcaat aagttccta cttggggag 
tgtttcaag tcacgaaat tgtaccaaa tatccaacc 
gatccaaat ttttgcgac aaaatcaaa ttatcggtc 
aatattcac aaagaactg accagaatt tgtagagct 
atcaactcg attttttca gtttttttt ctgctttgg 
gtcactgtg aatttcttg tgcttggtt tcagatctg 
tacattgtt gcttattta atctcaagc ggccagttt 
tttaattat tggcaaggc gagttgaaa gcttttaag 
aacgtttgt atttattcg attgagatg ttttatatt 
gcgagaaga tcgtcacgg ctttgttct caagcaaac 
agaactaaa actattttg gttgggttg aaagtggat 
ttaagcaaa gaggaggaa cgaaatact attattact 
tcgagtctt gaacttcga cagtttact cttcaaata 
acggcttgt agacttttt acgattgat aatgcgtta 
cttttttcg atctgtgga gctgccttt tcatacctc 
ttcataatg atacaaatg gacaaagac ttgaataag 
tctctccga aatgtaaca aattctatt ctatagttt 
gcgaaataa ataaacaat ctcatgcaa c
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY