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Gene information
General information
TC032947
Former TC numbers

If one gene is splitted or genes are merged, the original TC numbers are listed. One listed former TC number indicates the previous gene has be splitted, serveral former TC number represents a merged gene.

TC008393 		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_007419.2: 9656928..9661224
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
      		TC032947
      iBeetle screen iB_07243

      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: 3 (3 dpi)
      Days post injection

      adult lethality - Number of animals showing the phenotype: 1 (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
      • >TC032947 RA
        MAQCFVSTY RNHYRWPDP QFTKSAHEP 
YDNFVSVIP PEDIWIGAD TGEDKLDAA 
ALEKRLRKC YLKRLITTY QYDYCKDLV 
AKFIGEPAN IEIENQLLQ SAKISTILT 
PVRNRSPVT GTRKFLVIS DPVETTRP
      CDS sequence
      • >TC032947 RA
        atggcccaa tgtttcgtt tccacttat cgaaatcac 
tacaggtgg cccgacccc caattcaca aaatctgcc 
catgagccg tacgataac tttgtcagt gttattccc 
cctgaagac atttggatt ggcgccgat accggcgag 
gataaattg gacgctgca gctttggag aaacgcctg 
agaaaatgt tatctcaaa cgcctaata acaacctat 
cagtatgat tattgcaag gacttggtg gctaaattc 
atcggcgag cctgcaaac atcgaaatc gagaatcag 
ctcctgcaa tcggcaaaa atctctacg atcctgact 
cccgtccga aatcgctcc cccgttacg ggaacgaga 
aagtttttg gttatatcc gacccagtc gaaactaca 
cgcccttga
      mRNA sequence
      • >TC032947 RA
        aatttttga cagtaaaat tgacgtttt ttttggaaa 
attcacgtg aaaatggcc caatgtttc gtttccact 
tatcgaaat cactacagg tggcccgac ccccaattc 
acaaaatct gcccatgag ccgtacgat aactttgtc 
agtgttatt ccccctgaa gacatttgg attggcgcc 
gataccggc gaggataaa ttggacgct gcagctttg 
gagaaacgc ctgagaaaa tgttatctc aaacgccta 
ataacaacc tatcagtat gattattgc aaggacttg 
gtggctaaa ttcatcggc gagcctgca aacatcgaa 
atcgagaat cagctcctg caatcggca aaaatctct 
acgatcctg actcccgtc cgaaatcgc tcccccgtt 
acgggaacg agaaagttt ttggttata tccgaccca 
gtcgaaact acacgccct tgacgacct accagttct 
ttatcggaa aaacgggct atcaaatgc tcaaaggca 
tgccacagg ccgaacccc agcaacaag acgagaaag 
gggaaaatt tggtgaaaa aaccggaaa aaatcgaag 
cgaaatgcg aatgctatt gttgctcaa ggaaataaa 
aattcgcca ccacgtgta aaacaaatt ttattagtt 
gttttgagt cgttgtaaa tgcgctgtg aaaaataaa 
atatttccg aattt
      The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY