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Gene information
General information
TC003488 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: 5884024..5885762
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
    		TC003488
    iBeetle screen iB_06931

    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: 40.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 1 (3 dpi)
    Days post injection

    adult lethality - Number of animals showing the phenotype: 3 (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): more than 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    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): more than 80%
    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
    • >TC003488 RA
      MNAEVLRKS PNKNNKENE NGAEPSAPI SKKGILTSF 
LTGKSMEIP DSDILGRCR FVSEFEKLN RIGEGTYGI 
VYRAKDTIS DKIVALKKV RMDLERDGI PVSSLREIQ 
VLLKCRHEN IVHLKEVVV GRSLESIFL AMEYCEQDL 
ASLLDNMQA PFTESQVKC IMLQVLRGL RYLHHNFVV 
HRDLKVSNL LMTDKGCVK IADFGLARW FGVPLRPMT 
PHVVTLWYR APELLLQAP TQTTSVDMW AAGCILGEL 
LGHKPLLPG RSEIQQLEL IVDLLGTPS DAIWPGFSE 
LPALENFSL KQQPYNNLK QRFPWLSAA GLRLLNFLF 
MYDPRKRAT AEECLQSSY FKEPPLPCD PKLMPTFPQ 
HRNIKGGKT SAPSDSANT GAGDQTNNL PAISDLLGS 
LVKKRRIE
    CDS sequence
    • >TC003488 RA
      atgaacgct gaagtatta aggaaatcc cctaacaaa 
aacaataaa gaaaacgaa aatggggcc gaaccgtcg 
gcccctatt tcaaaaaag ggaatcttg acatcattc 
cttaccggc aaatcaatg gaaatacca gattccgac 
attttgggg cggtgccgg tttgtatca gagtttgag 
aagctgaac agaattggc gaaggcact tacggaata 
gtttaccgc gccaaggac acaatttcc gataaaatc 
gtggctctt aaaaaagtc cgcatggac ttggagcgg 
gacgggatt cccgtcagc agtttgcgc gaaattcag 
gttctcctg aaatgccgt cacgaaaat atcgtgcat 
ttgaaggag gttgttgtg gggcgcagc ttggagagc 
atttttctg gctatggag tattgcgag caagacctt 
gcctcgctg ttggataac atgcaagcc ccctttact 
gagtcccaa gtgaagtgt atcatgctg caagtgttg 
agaggactg cggtatttg caccacaat tttgtcgtg 
caccgcgat ttgaaagtc tccaatttg ctgatgacg 
gataagggc tgcgttaaa atcgccgat tttggttta 
gcgcgctgg tttggggtt cccttgcgt cccatgacc 
ccgcatgtg gtcacgctg tggtacagg gccccagag 
ttgctcctg caagcgccc acgcagact acgagtgtt 
gatatgtgg gctgcgggt tgtatttta ggggagctt 
ctgggccat aaacctttg ctacctgga aggtctgaa 
atacagcag cttgaactt attgtggac ttgcttggt 
accccatct gatgcgatt tggcctgga tttagtgaa 
ttaccagca ctggaaaat ttttctctg aaacagcag 
ccttacaat aacttaaaa cagagattt ccgtggctg 
tctgctgct ggtttgaga ctattaaac tttttgttt 
atgtatgat ccacgtaag cgcgctact gctgaagaa 
tgtttacaa agtagttat ttcaaggaa cctccacta 
ccttgtgac ccaaaattg atgccaaca ttcccccaa 
cacaggaat attaaaggc ggtaagaca tcggcgcct 
tcagatagt gcaaacact ggagctggt gaccaaact 
aataatttg ccggctata tcagatttg ttgggttcg 
cttgtgaag aagaggcgt attgaataa
    mRNA sequence
    • >TC003488 RA
      ttgcacgcc gtaaattac tagaattgt ttacggcgt 
tttttaatt tacttttat cgataatta ttaaaatac 
aataatttg attgtcgat tcaacgtaa aaatgaacg 
ctgaagtat taaggaaat cccctaaca aaaacaata 
aagaaaacg aaaatgggg ccgaaccgt cggccccta 
tttcaaaaa agggaatct tgacatcat tccttaccg 
gcaaatcaa tggaaatac cagattccg acattttgg 
ggcggtgcc ggtttgtat cagagtttg agaagctga 
acagaattg gcgaaggca cttacggaa tagtttacc 
gcgccaagg acacaattt ccgataaaa tcgtggctc 
ttaaaaaag tccgcatgg acttggagc gggacggga 
ttcccgtca gcagtttgc gcgaaattc aggttctcc 
tgaaatgcc gtcacgaaa atatcgtgc atttgaagg 
aggttgttg tggggcgca gcttggaga gcatttttc 
tggctatgg agtattgcg agcaagacc ttgcctcgc 
tgttggata acatgcaag cccccttta ctgagtccc 
aagtgaagt gtatcatgc tgcaagtgt tgagaggac 
tgcggtatt tgcaccaca attttgtcg tgcaccgcg 
atttgaaag tctccaatt tgctgatga cggataagg 
gctgcgtta aaatcgccg attttggtt tagcgcgct 
ggtttgggg ttcccttgc gtcccatga ccccgcatg 
tggtcacgc tgtggtaca gggccccag agttgctcc 
tgcaagcgc ccacgcaga ctacgagtg ttgatatgt 
gggctgcgg gttgtattt taggggagc ttctgggcc 
ataaacctt tgctacctg gaaggtctg aaatacagc 
agcttgaac ttattgtgg acttgcttg gtaccccat 
ctgatgcga tttggcctg gatttagtg aattaccag 
cactggaaa atttttctc tgaaacagc agccttaca 
ataacttaa aacagagat ttccgtggc tgtctgctg 
ctggtttga gactattaa actttttgt ttatgtatg 
atccacgta agcgcgcta ctgctgaag aatgtttac 
aaagtagtt atttcaagg aacctccac taccttgtg 
acccaaaat tgatgccaa cattccccc aacacagga 
atattaaag gcggtaaga catcggcgc cttcagata 
gtgcaaaca ctggagctg gtgaccaaa ctaataatt 
tgccggcta tatcagatt tgttgggtt cgcttgtga 
agaagaggc gtattgaat aaaggggtc aatttttga 
taaacaaat gttgtgttt tgtgtgcta tgataatgt 
agattgtgt ttattaatt attcataca caaataaac 
gtatttttt aagt
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY