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Gene information
General information
TC003556 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: 7269575..7270347
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
    		TC003556
    iBeetle screen iB_03577

    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)

    Fertility
    tropharium shortened - Number of animals showing the phenotype: 4

    tropharium structure irregular - Number of animals showing the phenotype: 4

    Comment: Tropharia seem to be filled with proocytes
    vitellogenic egg chamber mostlystage sequence not present - Number of animals showing the phenotype: 4

    previtellogenic egg chamber number decreased - Number of animals showing the phenotype: 4

    egg chamber stage sequence irregular - Number of animals showing the phenotype: 4
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 20
    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: 70.0% (includes death as larva, prepupa, pupa, adult)

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

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 5 (16 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 1 (19 dpi)
    Days post injection

    remarks: later than schedule,
    Pupal morphology
    antenna size shortened - Number of animals showing the phenotype: 5

    wings size shortened - Number of animals showing the phenotype: 5

    legs orientation irregular - Number of animals showing the phenotype: 5

    Adult morphology
    remarks: later than schedule,
    Fertility
    egg production: number no/almost no eggs -
    tropharium size decreased - Number of animals showing the phenotype: 3

    tropharium shape rounded - Number of animals showing the phenotype: 3
    egg chamber mostlynot present - Number of animals showing the phenotype: 3
    egg chamber mostlydeposition - Number of animals showing the phenotype: 3
    Comment: it seems that there is sometimes one egg-chamber depositioned in the tropharium
    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC003556 RA
      MSNLDRCFV SGWSDDEDG IDPERNPTE 
ENPKRILAA AENGELNEI KSLIELDPS 
LIHSTDKEG YTALHRACY GNHVEVVKY 
LLKNGADIA AKTEMQWQP LHSCCHWNN 
IECAQILLA EGADVNALS EGGQTPLHI 
AASHGARYD LVQLLLMHP YIKPDLKNN 
SNETAFDIA RRSSKYRNV FDMVDPLID 
LKHLEM
    CDS sequence
    • >TC003556 RA
      atgtccaat cttgaccgc tgttttgtc agtggttgg 
tcagacgat gaggatggc attgatccc gaacgcaat 
cccaccgaa gagaacccc aaacgcatt ttagcggcc 
gccgaaaat ggcgagttg aatgaaatc aagagctta 
atcgaactc gaccctagt ttaattcat tcaaccgat 
aaggaaggc tacactgct cttcacagg gcttgttat 
ggcaatcac gttgaagtc gtgaaatat ttgcttaaa 
aacggtgca gacatcgca gctaagacg gaaatgcag 
tggcagcct ttgcactct tgctgtcac tggaacaac 
atcgagtgt gcacagatt ctgctggcc gagggggct 
gatgtgaat gcgttgtct gaagggggg caaactccc 
ctgcatatt gccgcgtct catggggcg cgttacgac 
ctggtccag ttgctgcta atgcatccc tatattaag 
ccagatttg aagaacaac agcaatgag actgcgttt 
gatattgca aggcgcagc tcaaagtat cggaatgtt 
tttgacatg gtcgacccc ctaatagac ttaaaacac 
ttagaaatg taa
    mRNA sequence
    • >TC003556 RA
      ttagttgtc aacagcaac ataacctcg aataactga 
actcaagaa aaaataaat gcgaaatgt ccaatcttg 
accgctgtt ttgtcagtg gttggtcag acgatgagg 
atggcattg atcccgaac gcaatccca ccgaagaga 
accccaaac gcattttag cggccgccg aaaatggcg 
agttgaatg aaatcaaga gcttaatcg aactcgacc 
ctagtttaa ttcattcaa ccgataagg aaggctaca 
ctgctcttc acagggctt gttatggca atcacgttg 
aagtcgtga aatatttgc ttaaaaacg gtgcagaca 
tcgcagcta agacggaaa tgcagtggc agcctttgc 
actcttgct gtcactgga acaacatcg agtgtgcac 
agattctgc tggccgagg gggctgatg tgaatgcgt 
tgtctgaag gggggcaaa ctcccctgc atattgccg 
cgtctcatg gggcgcgtt acgacctgg tccagttgc 
tgctaatgc atccctata ttaagccag atttgaaga 
acaacagca atgagactg cgtttgata ttgcaaggc 
gcagctcaa agtatcgga atgtttttg acatggtcg 
accccctaa tagacttaa aacacttag aaatgtaat 
gatatgtta ttaatttat aattatgta ttttgttat 
ctttgtttt atatttttt gtaataaat aaaatcatt 
atta
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY