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Gene information
General information
TC005461 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_007423.3: 12758375..12759676
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
    		TC005461
    iBeetle screen iB_03957

    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: 0.0% (includes death as pupa, adult)

    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature irregular - Penetrance (percentage of animals/eggs on slide): more than 80%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - 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%
    larva complete everted (inside-out) - 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): 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: 30.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 50.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality - Number of animals showing the phenotype: 3 (11 dpi)
    Days post injection

    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 2 (16 dpi)
    Days post injection

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC005461 RA
      MGILEKISE IEKEISRTQ KNKATEYHL GLLKAKLAK 
YRSQLLEPS KKSEKGEGF DVLKSGDAR VALIGFPSV 
GKSTLLSTL TKTESEAAS YEFTTLTCI PGVIDYNGA 
NIQLLDLPG IIEGAAQGK GRGRQVIAV ARTADLVLM 
MLDATKKDV HRHLLEKEL ESVGIRLNK MKPNIYFKV 
KKGGGISFN STCPLTKID EKLVQMILH EYKIFNAEV 
LFREDCTAD ELIDVICAN RVYLPCLYV YNKIDQISI 
EEVDRIARQ PNSVVVSCN MKLNLDYLL QVLWEYLNL 
IRVYTKKPG QPPDFSDGL ILRKGVTVE HVCHAIHRT 
LAQQFKYAL VWGTSTKYS PQRVGTSHV MHDEDVIQI 
VKK
    CDS sequence
    • >TC005461 RA
      atgggaatt ttagaaaaa atctcggaa atcgagaag 
gaaatctct cgcacgcaa aaaaataaa gccaccgag 
taccactta ggtctcctc aaagccaaa cttgcaaaa 
taccgttcc cagctcttg gagccttcg aaaaaatcg 
gaaaagggt gaaggtttc gacgtttta aaaagtggg 
gatgcgcgt gtggccttg atcggcttc ccgtcagtc 
ggaaaatcc actttgttg tcaacttta acgaagacg 
gaaagcgag gccgcgtcg tacgagttc acgaccttg 
acgtgcatc ccgggcgtg atcgactac aatggggcc 
aatatccag ctgctcgat ttgcccggg attatcgag 
ggggctgcc cagggcaag ggcaggggg cggcaggtg 
atagcggtg gcgcgcacg gccgacttg gtcctcatg 
atgctggac gccacgaaa aaggacgtc caccgacac 
ttattagaa aaggaattg gaaagtgtg gggattagg 
ctgaacaaa atgaagccc aacatttat ttcaaggtg 
aaaaaaggg ggcggtatt tcgttcaat tccacttgt 
cccttgacg aaaatcgac gaaaagctc gtgcaaatg 
attctgcac gagtacaaa atcttcaac gctgaggtt 
ctcttcagg gaagactgt actgccgat gagttaatc 
gacgtgatt tgtgcgaac cgcgtctat cttccatgc 
ctttacgtt tataataaa atcgatcag atttcgatc 
gaggaagtc gatagaata gccagacag ccgaacagt 
gttgtcgtc agttgcaac atgaagctc aacctagat 
tacctcctt caagtcctc tgggagtat ttaaattta 
attcgcgtt tatacgaaa aaaccggga cagccgccg 
gatttcagc gacggcttg attctgaga aagggggtt 
acagtggag catgtgtgt cacgcaatc cacagaacg 
ctcgcccag caattcaag tatgcctta gtctggggc 
acgagcacc aaatactcg ccccaaagg gtcggcact 
tcccacgtc atgcacgac gaggacgtc atccaaata 
gttaagaag taa
    mRNA sequence
    • >TC005461 RA
      gaataaaac gtgccagaa aatgggaat tttagaaaa 
aatctcgga aatcgagaa ggaaatctc tcgcacgca 
aaaaaataa agccaccga gtaccactt aggtctcct 
caaagccaa acttgcaaa ataccgttc ccagctctt 
ggagccttc gaaaaaatc ggaaaaggg tgaaggttt 
cgacgtttt aaaaagtgg ggatgcgcg tgtggcctt 
gatcggctt cccgtcagt cggaaaatc cactttgtt 
gtcaacttt aacgaagac ggaaagcga ggccgcgtc 
gtacgagtt cacgacctt gacgtgcat cccgggcgt 
gatcgacta caatggggc caatatcca gctgctcga 
tttgcccgg gattatcga gggggctgc ccagggcaa 
gggcagggg gcggcaggt gatagcggt ggcgcgcac 
ggccgactt ggtcctcat gatgctgga cgccacgaa 
aaaggacgt ccaccgaca cttattaga aaaggaatt 
ggaaagtgt ggggattag gctgaacaa aatgaagcc 
caacattta tttcaaggt gaaaaaagg gggcggtat 
ttcgttcaa ttccacttg tcccttgac gaaaatcga 
cgaaaagct cgtgcaaat gattctgca cgagtacaa 
aatcttcaa cgctgaggt tctcttcag ggaagactg 
tactgccga tgagttaat cgacgtgat ttgtgcgaa 
ccgcgtcta tcttccatg cctttacgt ttataataa 
aatcgatca gatttcgat cgaggaagt cgatagaat 
agccagaca gccgaacag tgttgtcgt cagttgcaa 
catgaagct caacctaga ttacctcct tcaagtcct 
ctgggagta tttaaattt aattcgcgt ttatacgaa 
aaaaccggg acagccgcc ggatttcag cgacggctt 
gattctgag aaagggggt tacagtgga gcatgtgtg 
tcacgcaat ccacagaac gctcgccca gcaattcaa 
gtatgcctt agtctgggg cacgagcac caaatactc 
gccccaaag ggtcggcac ttcccacgt catgcacga 
cgaggacgt catccaaat agttaagaa gtaattaga 
ttcgttgtt actgcatat tattaaatg attttctta 
cac
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY