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Gene information
General information
TC004221 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_seqid2071: 596..2640
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
    TC004221
    iBeetle screen iB_03707
    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: 10.0% (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


    remarks: later than schedule, hours 48 remarks: later than schedule, hours 48 remarks: later than schedule, hours 48
    Pupal morphology
    remarks: later than schedule, hours 48
    Adult morphology
    remarks: later than schedule, hours 48 remarks: later than schedule, hours 48
    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 remarks: later than schedule, hours 48 (egglay)
    remarks: later than schedule, hours 48
    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%
    remarks: later than schedule, hours 48
    remarks - cuticles: later than schedule, hours 48
    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: 10.0% (includes death as larva, prepupa, pupa, adult)

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


    adult lethality - Number of animals showing the phenotype: 2 (41 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC004221 RA
      MLCEAHLVN SLTFLSKVL HTHYEEKVF ILIDEYDAV 
      VGLVFESCP NKDDIGLIN DCLKRFMAN TLKFNDFIK 
      RSLIFACVR LAGAFSGDA ARVIEHYPF LSDLRYAPY 
      LGFTDEEVK HLLELPEVK AACGKVTFK TITEWYDGY 
      MVSNSPLKI FSSWSVLNL LKRVYTTGT LKYSSFWII 
      NPGHVKNLG KLFAEPSVR EKIEELISN GEVKIGRVT 
      SVNVQHLKS LQQLISLEF DEISEENVE LFLHFLADS 
      GYFNILSID KKTDDIETN TWDEDEDED EDEDEDEEW 
      ADFESDSTF CVLKIPNFE IKQHVRNLC YSEALAVIR 
      KFHLNARSV KNYVSALNI LLQSTSERA FVDLANAVL 
      VLLSESQKI MNEKELHHI LYTLVYDSR YFETHSEVQ 
      IKKKEKDQG
    CDS sequence
    • >TC004221 RA
      atgttatgt gaagctcat ttggtaaat agcttaaca 
      tttttatcg aaagtttta catacacat tatgaggaa 
      aaagttttc atactcatc gatgagtac gatgctgtg 
      gtgggcctg gtattcgag tcttgtccg aataaagat 
      gacattgga ttaattaat gattgtcta aaacgtttc 
      atggctaat acgctgaaa ttcaatgat ttcataaaa 
      cgctcatta atttttgct tgtgttcgg ttggccgga 
      gccttctca ggggatgca gcaagagtt atagaacat 
      tatccgttc ctaagtgat cttcgatac gcaccgtac 
      ttaggattt acggatgag gaggtaaag catctactt 
      gagcttcca gaagttaaa gctgcttgt gggaaagta 
      acattcaaa acaattacg gaatggtat gatggatat 
      atggtatct aatagccca ttgaaaata ttcagtagc 
      tggtcagtc ttgaacctt ttgaaaagg gtttataca 
      accggtacg ctcaaatat tcctctttc tggattatc 
      aatccaggt catgtaaaa aatctgggg aagctgttt 
      gcggaacct tcagttcga gaaaaaatt gaggaactt 
      atttctaat ggtgaagtt aagatagga agagttaca 
      tcagtcaat gtccaacat ctaaagagt ttacagcaa 
      ctgatttct ttagagttt gatgaaata tcggaggaa 
      aatgtggag ttatttttg catttcttg gctgattct 
      ggttacttt aatatattg tcaattgat aagaaaaca 
      gatgacatt gaaacaaac acctgggat gaagatgaa 
      gatgaagat gaagatgaa gatgaagat gaagaatgg 
      gctgatttt gaaagtgac tcgacattt tgtgtttta 
      aaaatacct aattttgaa attaagcaa catgtccgc 
      aacctttgc tatagtgaa gcactggca gttatacgt 
      aaattccac ttgaacgct agatctgtt aaaaattat 
      gtaagtgct ctgaatata ctattacaa agtacaagt 
      gagagggca tttgttgat cttgcaaac gccgtatta 
      gtattactc tctgaatca cagaagatt atgaacgaa 
      aaagaactt catcacatc ttgtatact ctggtatat 
      gactccaga tacttcgaa acacacagt gaggtccag 
      atcaagaaa aaagaaaag gaccaggga tag
    mRNA sequence
    • >TC004221 RA
      ttgtattac agattgtag ccgaaccaa agtaaaaat 
      gaacgagag ttcttctct aaatgattc taaggatca 
      gaaaagtcg ctgtgcgaa ctggcaagt catgaattc 
      gccacaatc aaagagaca attaccttt gtggacaag 
      accttgttt atcaaatgt ctcttaagt gatactagt 
      gaaatagta ttactaacg gcgcctaga gcttttgga 
      aaaactacc ctcatggat atgttgaag cagtttcta 
      ctgggacga aaaagttta tttgagaac ctaaagatt 
      tgtactgaa gaaacacaa tttttttca aagcactgc 
      agatctcat cctgtgata catattgat ctcgggagt 
      gttcgtggt agcagtttt actgaagtt aagcacgaa 
      ttggcgttg gctataaat cgtgcattc agagaacat 
      gaatatctg gtgaagaag actgatgat ggtagaata 
      gtctggaca agtgaaaag ttaagcatt tctaactta 
      aaaatagca agttttgaa aatttctac gttcaaaaa 
      aagtgcatc atgttatgt gaagctcat ttggtaaat 
      agcttaaca tttttatcg aaagtttta catacacat 
      tatgaggaa aaagttttc atactcatc gatgagtac 
      gatgctgtg gtgggcctg gtattcgag tcttgtccg 
      aataaagat gacattgga ttaattaat gattgtcta 
      aaacgtttc atggctaat acgctgaaa ttcaatgat 
      ttcataaaa cgctcatta atttttgct tgtgttcgg 
      ttggccgga gccttctca ggggatgca gcaagagtt 
      atagaacat tatccgttc ctaagtgat cttcgatac 
      gcaccgtac ttaggattt acggatgag gaggtaaag 
      catctactt gagcttcca gaagttaaa gctgcttgt 
      gggaaagta acattcaaa acaattacg gaatggtat 
      gatggatat atggtatct aatagccca ttgaaaata 
      ttcagtagc tggtcagtc ttgaacctt ttgaaaagg 
      gtttataca accggtacg ctcaaatat tcctctttc 
      tggattatc aatccaggt catgtaaaa aatctgggg 
      aagctgttt gcggaacct tcagttcga gaaaaaatt 
      gaggaactt atttctaat ggtgaagtt aagatagga 
      agagttaca tcagtcaat gtccaacat ctaaagagt 
      ttacagcaa ctgatttct ttagagttt gatgaaata 
      tcggaggaa aatgtggag ttatttttg catttcttg 
      gctgattct ggttacttt aatatattg tcaattgat 
      aagaaaaca gatgacatt gaaacaaac acctgggat 
      gaagatgaa gatgaagat gaagatgaa gatgaagat 
      gaagaatgg gctgatttt gaaagtgac tcgacattt 
      tgtgtttta aaaatacct aattttgaa attaagcaa 
      catgtccgc aacctttgc tatagtgaa gcactggca 
      gttatacgt aaattccac ttgaacgct agatctgtt 
      aaaaattat gtaagtgct ctgaatata ctattacaa 
      agtacaagt gagagggca tttgttgat cttgcaaac 
      gccgtatta gtattactc tctgaatca cagaagatt 
      atgaacgaa aaagaactt catcacatc ttgtatact 
      ctggtatat gactccaga tacttcgaa acacacagt 
      gaggtccag atcaagaaa aaagaaaag gaccaggga 
      tagccaata ccctagatg tggtgatat cagtaagta 
      cgttgga