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Gene information
General information
TC032664
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.

TC003974,TC003975 		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_007418.3: 29548724..29576877
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
    		TC032664
    iBeetle screen iB_03662

    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: 30.0% (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

    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: < 50 embryo/egg with embryonic tissue, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 50
     egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 50% - 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: 0.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 10.0% (includes death as larva, prepupa, pupa, adult)

    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (16 dpi)
    Days post injection

    adult lethality - Number of animals showing the phenotype: 4 (41 dpi)
    Days post injection

    Adult morphology
    elytra dorsal orientation not closed - Number of animals showing the phenotype: 3


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC032664 RA
      MLKERESIE RAISSPCSS LGGSSTLDG TTCAGCGGR 
IQDRYYLLA VDRQWHASC LKCCECKLP LDTELTCFA 
RDGNIYCKE DYYRMFAVT RCGRCQAGI SANELVMRA 
RDSVYHLHC FSCTSCGIP LSKGDHFGM RDGLIYCRP 
HYELLDFCD TSDPVEMMF RGSESPPGY FANAAPQHH 
KGRPRKRKL PVEDRGCGE IPVTMRMAA ATLEMLQQG 
DLSSSMESL SYDSSVTSP ASSNGHQPQ RTKRMRTSF 
KHHQLRTMK TYFAINQNP DAKDLKQLA QKTGLSKRV 
LQVWFQNAR AKWRRNIMR QEGGQSQGQ SQANGPPSV 
GSGIMGEGN SLSHQSLDD LHHHLHSQN SQTLTFSDI 
Y
    CDS sequence
    • >TC032664 RA
      atgctgaag gagcgcgaa tccatcgag cgtgccatc 
tcctcgccc tgctccagc cttggcggc tcctccacg 
ctggacggg acgacctgt gcgggctgc ggaggccga 
atccaggac cgttactac ctcctggcc gtcgacaga 
caatggcat gcctcctgc ttgaaatgt tgcgaatgt 
aaacttcct ctggatacg gagctgact tgtttcgcc 
cgagatggg aacatttac tgcaaggaa gattattat 
cggatgttt gcggtgacg cgctgtggc cgctgccag 
gcgggcatc tccgccaac gagctggtg atgcgagcc 
agagactcc gtctaccac ctgcactgc ttctcgtgc 
acctcctgc gggatcccg ctgtccaaa ggagaccac 
ttcgggatg agagatgga ctcatttat tgtcggcca 
cattacgag ctgctggat ttttgtgat acgagcgat 
ccggtggag atgatgttc agagggagc gagtcgcca 
ccggggtat tttgccaac gcggcgcca cagcaccac 
aaggggaga cccaggaag cggaaactg ccggtggag 
gatcggggg tgtggggaa attccggtc acaatgaga 
atggcagcg gcaacacta gagatgctc cagcaaggc 
gacctctcc tcgtccatg gagtcgctc tcgtacgac 
tcgtcggtg acgtccccc gcgtccagc aacggccac 
cagccccag cggaccaag cgcatgcgc acgtccttc 
aagcaccac cagctgagg accatgaag acgtacttc 
gccatcaat cagaacccc gacgccaag gacctgaag 
cagctggcg cagaagacg ggcctctcg aagagggtg 
ctccaggtg tggttccag aacgccagg gccaagtgg 
aggaggaac atcatgcgg caggaaggc ggccaaagc 
cagggccag agccaggcc aatgggccc ccgtccgtg 
ggctcgggg atcatgggg gaggggaac agtttgtcg 
catcagtcg ctagatgat ctccaccat catctgcat 
tcgcaaaat tcgcaaact ttgacgttc agtgatatt 
tattag
    mRNA sequence
    • >TC032664 RA
      cgacggtaa actgcacgg tggctgggc tcgaaacgt 
cgcgagtgt tgctgtgtt gtgctgttt tgacagttc 
ttgtggatt atgggcggc gttgccagg tagctgcag 
cttcagtcg tttttgggc tcgtgcaag atgctgaag 
gagcgcgaa tccatcgag cgtgccatc tcctcgccc 
tgctccagc cttggcggc tcctccacg ctggacggg 
acgacctgt gcgggctgc ggaggccga atccaggac 
cgttactac ctcctggcc gtcgacaga caatggcat 
gcctcctgc ttgaaatgt tgcgaatgt aaacttcct 
ctggatacg gagctgact tgtttcgcc cgagatggg 
aacatttac tgcaaggaa gattattat cggatgttt 
gcggtgacg cgctgtggc cgctgccag gcgggcatc 
tccgccaac gagctggtg atgcgagcc agagactcc 
gtctaccac ctgcactgc ttctcgtgc acctcctgc 
gggatcccg ctgtccaaa ggagaccac ttcgggatg 
agagatgga ctcatttat tgtcggcca cattacgag 
ctgctggat ttttgtgat acgagcgat ccggtggag 
atgatgttc agagggagc gagtcgcca ccggggtat 
tttgccaac gcggcgcca cagcaccac aaggggaga 
cccaggaag cggaaactg ccggtggag gatcggggg 
tgtggggaa attccggtc acaatgaga atggcagcg 
gcaacacta gagatgctc cagcaaggc gacctctcc 
tcgtccatg gagtcgctc tcgtacgac tcgtcggtg 
acgtccccc gcgtccagc aacggccac cagccccag 
cggaccaag cgcatgcgc acgtccttc aagcaccac 
cagctgagg accatgaag acgtacttc gccatcaat 
cagaacccc gacgccaag gacctgaag cagctggcg 
cagaagacg ggcctctcg aagagggtg ctccaggtg 
tggttccag aacgccagg gccaagtgg aggaggaac 
atcatgcgg caggaaggc ggccaaagc cagggccag 
agccaggcc aatgggccc ccgtccgtg ggctcgggg 
atcatgggg gaggggaac agtttgtcg catcagtcg 
ctagatgat ctccaccat catctgcat tcgcaaaat 
tcgcaaact ttgacgttc agtgatatt tattagggt 
agtgacact gtatagact agttaggtt cacacacgc 
aatgttcct tcatgatag tgataattg gatgatttt 
atcggttgt ttttcagtt gaggatttt gttagagga 
tagattgca cgatgagtt ttgaggatc ttgtagata 
atgtcgttg tttgtacat aagaatata cgttagtgg 
taggcgaga gttttgagg aatatttat aattttgta 
tgggtgtgc gatgatgtc atacatgtg cggtgtatg 
tctacaagg aataaatgt atataattt at
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY