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Gene information
General information
TC004216 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_seqid1168: 586..3060
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
    TC004216
    iBeetle screen iB_03706
    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 embryo/egg not developed/not fertilized - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    embryo/egg with embryonic tissue, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    larval musculature irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    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
    egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): more than 80%

    Comment: > 100 empty eggs on slide
    cuticle remnants with some identifiable segments - Penetrance (percentage of animals/eggs on slide): less than 30%

    Comment: only 5 eggs with remnants on the slide
    appendages partially remaining - Penetrance (percentage of animals/eggs on slide): less than 30%
    abdomen partially # of remaining segments remaining - Penetrance (percentage of animals/eggs on slide): less than 30%
    larva everted (inside-out) - Penetrance (percentage of animals/eggs on slide): less than 30%

    Comment: only 4 larvae inside-outed on the slide
    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: 70.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 80.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality - Number of animals showing the phenotype: 6 (11 dpi)
    Days post injection


    pupal molt not fulfilled (died as prepupa) - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection



    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: 1 (41 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC004216 RA
      MYISEFLVV FALCAFGIN ADPTCPKQD GKDSVYFPH 
      EDCTKFWQC SNGTPYLFD CPDNLHFNP KLNVCDWPN 
      AAGCKGSGE DSDSSSSSS SSSSSESQE SGDNSQGKD 
      DNNSSSSSS SSSSSSSSE EGSSPECPS VDGEDPVYF 
      PHEDCTKFW QCSNGVPYL FNCSANLHF NPKLNVCDW 
      PDQAGCESK EDSSSGSES KESDDKDDS SSSSSSSSS 
      SESQESGDN SQGKDDNNS SSSSSSSSS SSSEEGSSP 
      ECPSVDGED PVYFPHEDC TKFWQCSNG VPYLFNCSA 
      NLHFNPKLN VCDWPDQAG CESKEDSSS GSESKESDD 
      KDDSSSSSS SSSSSESKE SGDNSESCT SSSEEGPEC 
      PSVDGEDPV YFPHEDCTK FWQCSNGVP YLFNCSANL 
      HFNPKLNVC DWPDQAGCE SKEDSSSSS SSSSSESGD 
      NSQGKDDDK DDSGNSSSS SSDSSSSSS SSESSEEGP 
      ECPSVDGET PVYIPHEDC TKFWQCSNG TPYLFDCPD 
      NLHFNPKLN VCDWPNAAG CKGNK
    CDS sequence
    • >TC004216 RA
      atgtacatt tcagaattt ttagttgta tttgcctta 
      tgcgctttt ggaataaac gcagatcca acttgtccg 
      aaacaagat ggaaaagat tccgtctac tttccacat 
      gaagactgc accaaattt tggcagtgt tcaaatggc 
      actccgtat ctttttgac tgccctgac aatttgcat 
      tttaatccg aaactgaac gtttgtgat tggccaaat 
      gctgccgga tgcaaagga agcggtgaa gatagtgac 
      agcagtagc agtagcagc agcagcagt agtagcgaa 
      agtcaagaa tccggagat aactcacaa ggaaaagac 
      gataataac agtagcagt agcagtagt agcagcagc 
      agcagcagc agcagtgaa gaaggcagc agtcctgaa 
      tgtccctca gtagatgga gaagatcct gtctacttc 
      ccacatgaa gattgtaca aaattttgg caatgttct 
      aatggtgtt ccctacttg ttcaactgc tcagctaat 
      ttgcatttc aatcctaaa ttaaacgtt tgcgattgg 
      cccgatcaa gcaggttgt gaaagcaaa gaagatagt 
      agcagtggc agcgaaagt aaagaatct gatgataaa 
      gatgacagc agtagcagt agcagcagc agcagtagt 
      agcgaaagt caagaatcc ggagataac tcacaagga 
      aaagacgat aataacagt agcagtagc agtagtagc 
      agcagcagc agcagcagt gaagaaggc agcagtcct 
      gaatgtccc tcagtagat ggcgaagat cctgtctac 
      ttcccacat gaagattgt acaaaattt tggcaatgt 
      tctaatggt gttccctac ttgttcaac tgctcagct 
      aatttgcat ttcaatcct aaattaaac gtttgcgat 
      tggcccgat caagcagga tgtgaaagc aaagaagat 
      agtagcagt ggcagcgaa agtaaagaa tctgatgat 
      aaagatgac agcagcagc agtagcagc agcagcagc 
      agtagcgaa agcaaagaa tctggcgac aattccgaa 
      agctgcacc agcagcagt gaagaaggt cctgaatgt 
      ccttcagta gatggagaa gatcccgtc tacttccca 
      cacgaagat tgtaccaaa ttttggcaa tgttctaat 
      ggtgttccc tacttgttc aactgctca gctaatttg 
      catttcaat cctaaatta aacgtttgc gattggccc 
      gatcaagca ggttgtgaa agcaaagaa gatagcagc 
      agcagcagt agtagtagc agcagtgaa agcggtgac 
      aattcccaa ggaaaagac gacgataaa gatgatagt 
      ggtaacagc agcagcagc agtagtgac agtagcagt 
      agcagcagc agcagcgaa agtagtgaa gaaggtcct 
      gagtgtccc tcagtggac ggtgaaact ccagtttat 
      ataccacat gaagactgc accaaattc tggcaatgt 
      tcaaatggt actccgtat ctttttgac tgtccagac 
      aatttgcac tttaatccc aaattgaat gtttgtgat 
      tggccaaac gccgcagga tgtaaagga aacaaataa
    mRNA sequence
    • >TC004216 RA
      agttttgtg atttaacag tactaaacc tccttataa 
      ttatgtaca tttcagaat ttttagttg tatttgcct 
      tatgcgctt ttggaataa acgcagatc caacttgtc 
      cgaaacaag atggaaaag attccgtct actttccac 
      atgaagact gcaccaaat tttggcagt gttcaaatg 
      gcactccgt atctttttg actgccctg acaatttgc 
      attttaatc cgaaactga acgtttgtg attggccaa 
      atgctgccg gatgcaaag gaagcggtg aagatagtg 
      acagcagta gcagtagca gcagcagca gtagtagcg 
      aaagtcaag aatccggag ataactcac aaggaaaag 
      acgataata acagtagca gtagcagta gtagcagca 
      gcagcagca gcagcagtg aagaaggca gcagtcctg 
      aatgtccct cagtagatg gagaagatc ctgtctact 
      tcccacatg aagattgta caaaatttt ggcaatgtt 
      ctaatggtg ttccctact tgttcaact gctcagcta 
      atttgcatt tcaatccta aattaaacg tttgcgatt 
      ggcccgatc aagcaggtt gtgaaagca aagaagata 
      gtagcagtg gcagcgaaa gtaaagaat ctgatgata 
      aagatgaca gcagtagca gtagcagca gcagcagta 
      gtagcgaaa gtcaagaat ccggagata actcacaag 
      gaaaagacg ataataaca gtagcagta gcagtagta 
      gcagcagca gcagcagca gtgaagaag gcagcagtc 
      ctgaatgtc cctcagtag atggcgaag atcctgtct 
      acttcccac atgaagatt gtacaaaat tttggcaat 
      gttctaatg gtgttccct acttgttca actgctcag 
      ctaatttgc atttcaatc ctaaattaa acgtttgcg 
      attggcccg atcaagcag gatgtgaaa gcaaagaag 
      atagtagca gtggcagcg aaagtaaag aatctgatg 
      ataaagatg acagcagca gcagtagca gcagcagca 
      gcagtagcg aaagcaaag aatctggcg acaattccg 
      aaagctgca ccagcagca gtgaagaag gtcctgaat 
      gtccttcag tagatggag aagatcccg tctacttcc 
      cacacgaag attgtacca aattttggc aatgttcta 
      atggtgttc cctacttgt tcaactgct cagctaatt 
      tgcatttca atcctaaat taaacgttt gcgattggc 
      ccgatcaag caggttgtg aaagcaaag aagatagca 
      gcagcagca gtagtagta gcagcagtg aaagcggtg 
      acaattccc aaggaaaag acgacgata aagatgata 
      gtggtaaca gcagcagca gcagtagtg acagtagca 
      gtagcagca gcagcagcg aaagtagtg aagaaggtc 
      ctgagtgtc cctcagtgg acggtgaaa ctccagttt 
      atataccac atgaagact gcaccaaat tctggcaat 
      gttcaaatg gtactccgt atctttttg actgtccag 
      acaatttgc actttaatc ccaaattga atgtttgtg 
      attggccaa acgccgcag gatgtaaag gaaacaaat 
      aaattgtac agcacatac caaataatt acaaaatgt 
      taattacta ttctaaaat aaactgaaa aataaaac