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Gene information
General information
TC033445
Former TC numbers

If one gene is splitted or genes are merged, the original TC numbers are listed. One listed former TC number indicates the previous gene has be splitted, serveral former TC number represents a merged gene.

TC014925,TC014926 		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.

scaffold_seqid43: 2747504..2750154
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
    		TC033445
    iBeetle screen iB_02353
    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)

    adult lethality - Number of animals showing the phenotype: 4 (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): more than 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    remarks: muscle pattern potentially obscured by segmentation defect,
    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: 100.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality early (L5/6) - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection

    eclosion delayed (still pupa) - Number of animals showing the phenotype: 4 (16 dpi)
    Days post injection

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC033445 RA
      MGENVENTP GRQAIAVMK DIQKEIIFK KPVGTPRKR 
VKEKVLDED TYVEHIGKI IQRDFFPDL EKLKAQNEY 
LEAVERNDV TKMRQLYMK YSGNRPPTQ RSKCYKNKN 
YIMYIPDGV PLTQEEEME LNKNRQEVV HCNTRLTIN 
PFNEIQSKE AITELAKNQ AKIHDGKIG VDGKELMKD 
SPQIRGFSF VKTPSPHPN LTDTPLMTW GEIEGTPFR 
LDGSDTPLP HSQGPSFKM SEPPRREQL AIALAEKVG 
EKNRDRKQK AMEAARRRF ATPSPRTER LATMSPAAR 
RFATSRLGL RGDFMTPSP KIRATPGTP KRMVTPKLN 
LGIKKIDNL TDDLLKISV PKRHKAADF F
    CDS sequence
    • >TC033445 RA
      atgggtgaa aacgtggaa aatacaccc gggcggcag 
gcaattgcc gtgatgaaa gacatccag aaggaaatt 
attttcaaa aaaccggtg ggaacgcct cggaaacga 
gtcaaagag aaggtttta gatgaagac acttatgtg 
gagcacatt ggtaagatt atccagcgc gactttttc 
cccgatctt gaaaaactc aaagcccag aacgaatat 
ttggaagcc gttgagaga aacgatgtg accaaaatg 
aggcaactt tatatgaaa tatagcggg aatagaccg 
ccaacacag aggagtaag tgctacaaa aataaaaac 
tacataatg tacattcct gacggggtt cccctgaca 
caagaagaa gaaatggaa ttgaacaaa aaccggcaa 
gaagtcgtc cactgtaac acccgcctc accatcaac 
cctttcaac gaaatccaa agcaaggag gctatcacc 
gaactggcc aaaaaccag gcgaaaatc cacgacggt 
aaaatcggc gttgacggc aaagaactg atgaaagac 
agcccccag atcaggggc ttctccttc gtgaagacc 
ccctcgccc cacccgaac ctcaccgac acccccttg 
atgacgtgg ggcgaaatc gagggaacc ccctttcga 
ctggacggc agcgatacg cccctgccc cattcgcaa 
ggcccctcg tttaaaatg tcggagccc ccacgtcgt 
gaacagctc gccatagct ttggctgag aaagtcgga 
gagaagaat cgagacagg aaacagaag gctatggag 
gcggcgcga aggcggttt gcgacgccg tccccgagg 
acggaaaga ttggcaacc atgtcgccg gcggcgaga 
cgatttgcc acctccagg ttgggttta aggggtgat 
tttatgacg cccagtccg aaaataagg gccactccg 
gggactccc aagaggatg gttacgcct aaactgaat 
ctggggatc aagaaaatc gataatttg accgatgat 
ttgttgaag attagtgtg ccgaagagg cacaaagcc 
gccgatttt ttctaa
    mRNA sequence
    • >TC033445 RA
      atcacttaa aaatgggtg aaaacgtgg aaaatacac 
ccgggcggc aggcaattg ccgtgatga aagacatcc 
agaaggaaa ttattttca aaaaaccgg tgggaacgc 
ctcggaaac gagtcaaag agaaggttt tagatgaag 
acacttatg tggagcaca ttggtaaga ttatccagc 
gcgactttt tccccgatc ttgaaaaac tcaaagccc 
agaacgaat atttggaag ccgttgaga gaaacgatg 
tgaccaaaa tgaggcaac tttatatga aatatagcg 
ggaatagac cgccaacac agaggagta agtgctaca 
aaaataaaa actacataa tgtacattc ctgacgggg 
ttcccctga cacaagaag aagaaatgg aattgaaca 
aaaaccggc aagaagtcg tccactgta acacccgcc 
tcaccatca accctttca acgaaatcc aaagcaagg 
aggctatca ccgaactgg ccaaaaacc aggcgaaaa 
tccacgacg gtaaaatcg gcgttgacg gcaaagaac 
tgatgaaag acagccccc agatcaggg gcttctcct 
tcgtgaaga ccccctcgc cccacccga acctcaccg 
acaccccct tgatgacgt ggggcgaaa tcgagggaa 
ccccctttc gactggacg gcagcgata cgcccctgc 
cccattcgc aaggcccct cgtttaaaa tgtcggagc 
ccccacgtc gtgaacagc tcgccatag ctttggctg 
agaaagtcg gagagaaga atcgagaca ggaaacaga 
aggctatgg aggcggcgc gaaggcggt ttgcgacgc 
cgtccccga ggacggaaa gattggcaa ccatgtcgc 
cggcggcga gacgatttg ccacctcca ggttgggtt 
taaggggtg attttatga cgcccagtc cgaaaataa 
gggccactc cggggactc ccaagagga tggttacgc 
ctaaactga atctgggga tcaagaaaa tcgataatt 
tgaccgatg atttgttga agattagtg tgccgaaga 
ggcacaaag ccgccgatt ttttctaat tggagattg 
tttattttt ggttaataa atatttgtt aaaact