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Gene information
General information
TC004421 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_seqid6: 3313533..3314742
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
      TC004421
      iBeetle screen iB_03753
      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


      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): less than 30%

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

      larval musculature pattern 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): 30% - 50%
      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): 30% - 50%
      cuticle remnants without visible segmentation, differentiation or axis - Penetrance (percentage of animals/eggs on slide): less than 30%

      head mostlyremaining - 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): 50% - 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: 10.0% (includes death as larva, prepupa, pupa)
      Lethalities 22 days after larval injection: 30.0% (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: 1 (16 dpi)
      Days post injection


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


      RNAi sequence
      Primers
      Left: Right:
      Protein sequence
      • >TC004421 RA
        MGNEDTGEC SQQGDNCEF GRPLKKAKY VWQVKGKYH 
        LKASYQNKS EPSVDEDEE KRPCECAQH KASEERKCI 
        DNFLAKSES FFDSDEEEE VSKNIDKSI SNEIPITLV 
        SPYPKNQDY FLRKWQARQ IARGYIENT INSVLETWG 
        SAPFDAEDF VENCDEDGH IEDEGILVA IQAHGLQSN 
        LPKKENSLS ELAENALEK IDTVRSQIL RQELGLEPP 
        PECNPTKDC ESEDLSVEY NVVDILNSA ILVAIEKKG 
        LSSNND
      CDS sequence
      • >TC004421 RA
        atgggaaac gaagatact ggggagtgt tcgcaacag 
        ggcgataat tgcgagttt ggtaggccg ttaaagaaa 
        gccaaatac gtgtggcaa gtgaagggg aaataccac 
        cttaaagct agttaccag aacaaatcg gagccttct 
        gtggatgaa gatgaggag aaacgtcct tgtgaatgc 
        gctcagcat aaggcatca gaagagaga aaatgtatt 
        gataatttc cttgccaaa tctgagtcg ttctttgac 
        agtgatgag gaagaagaa gtctcaaaa aacattgac 
        aaatccata tcaaatgaa atacctatt actttagta 
        tcaccgtat cccaaaaac caggactat tttttgcga 
        aaatggcaa gcgagacaa atcgcgcgt ggttacatt 
        gaaaacacg ataaacagc gttttggag acttggggc 
        tctgccccc ttcgatgct gaagatttt gtcgaaaat 
        tgcgatgaa gatggacat attgaagat gaaggaatt 
        ttagtggca attcaagca catgggcta caatccaac 
        ttgccaaaa aaagagaac tctttgtct gaactcgca 
        gaaaatgct ttagaaaag attgacaca gtcaggagt 
        caaattctg agacaggag ttaggtctg gaaccacct 
        ccagagtgc aatccgaca aaagattgt gaatctgaa 
        gacttgagt gttgaatac aatgtggtg gatatcttg 
        aattctgca attctagtt gccattgaa aaaaagggg 
        ctgtcgtct aacaatgat taa
      mRNA sequence
      • >TC004421 RA
        acgtattaa acagtgacg attatgtgt ttgtgtgtc 
        ccaaaatag aaaaaaatt cctgtcatc tgtttttat 
        aattaaacg tcaaattga gccaaacaa ataagtgat 
        ttcagttcg atttcaaat gagaattgt tttcatttc 
        ttcttaaaa atgaattaa ccttttgga tgaacgtac 
        aaagccgtc gttttccta ccatttcga acgcgtaac 
        gtaaatttt gcgggatca atacttggg ctagttgca 
        atgggaaac gaagatact ggggagtgt tcgcaacag 
        ggcgataat tgcgagttt ggtaggccg ttaaagaaa 
        gccaaatac gtgtggcaa gtgaagggg aaataccac 
        cttaaagct agttaccag aacaaatcg gagccttct 
        gtggatgaa gatgaggag aaacgtcct tgtgaatgc 
        gctcagcat aaggcatca gaagagaga aaatgtatt 
        gataatttc cttgccaaa tctgagtcg ttctttgac 
        agtgatgag gaagaagaa gtctcaaaa aacattgac 
        aaatccata tcaaatgaa atacctatt actttagta 
        tcaccgtat cccaaaaac caggactat tttttgcga 
        aaatggcaa gcgagacaa atcgcgcgt ggttacatt 
        gaaaacacg ataaacagc gttttggag acttggggc 
        tctgccccc ttcgatgct gaagatttt gtcgaaaat 
        tgcgatgaa gatggacat attgaagat gaaggaatt 
        ttagtggca attcaagca catgggcta caatccaac 
        ttgccaaaa aaagagaac tctttgtct gaactcgca 
        gaaaatgct ttagaaaag attgacaca gtcaggagt 
        caaattctg agacaggag ttaggtctg gaaccacct 
        ccagagtgc aatccgaca aaagattgt gaatctgaa 
        gacttgagt gttgaatac aatgtggtg gatatcttg 
        aattctgca attctagtt gccattgaa aaaaagggg 
        ctgtcgtct aacaatgat taaactcga gcaaagact 
        gtttttttc tattataaa ttttgtatg tatttgtaa 
        gaattattt tgtgttatt actagaatt tttatttcc 
        tcattttat catcacatg aataaaagt accttgaaa 
        gt