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Gene information
General information
TC016225 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_007425.3: 4483071..4486863
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
    		TC016225
    iBeetle screen iB_05978

    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: 20.0% (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: 1 (9 dpi)
    Days post injection

    Fertility
    eggs lost during procedure: potentially due to defects in egg-shells egg-morphology or egg-size, (14 dpi)
    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: < 20 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
    • >TC016225 RA
      MATVKQVGD YKLGALIIE GKTKQVYEL PGSPDLCLL 
LNKDRITAG DGVKAHDLA GKAAISNKT NGKVFEILN 
SVGVKTSFV KAASDVAFI SKKCEMIPI EWVTRRLAT 
GSFLKRNPG VPEGYRFFP PKHETFFKD DANHDPQWS 
SEQIISAKF KVNGLEIGP TEVDIMTRM SILVFEILE 
KVWASRNCA LIDMKIEFG VDKSGEILV ADVIDSDSW 
RLWPSGDKR LMVDKQVYR NLTTVTSSD LDTVKRNFE 
WIVDQLDHL APPNDHLVV ILMGSPSDA DHCKTIQKH 
CEKLGLNTE LRVTSAHKG TEETIKIIR YYESLPLKV 
VFIAVAGRS NGLGPVVSG NSPFPVINC PPLKAENVD 
RDVWSSLNV PSGLGCSTV LYPEAAALS AANIIGLGN 
YVVWSRLRV RQLDNYVTL AKADKGVKN N
    CDS sequence
    • >TC016225 RA
      atggcgacg gtcaaacag gtcggtgac tacaaattg 
ggggccctc atcatcgag ggcaagacc aagcaagtg 
tacgagttg ccgggatcc cccgatttg tgccttttg 
ctgaataag gaccggatc acggcaggc gatggcgtc 
aaagcgcac gatctggcg ggaaaggcg gcaatttcg 
aataagaca aatggaaaa gttttcgaa attttaaat 
tcagtcggg gtgaaaact tcgttcgtt aaagccgcc 
tctgatgtg gcttttatt tcgaaaaag tgtgagatg 
atccccatt gagtgggtg acaaggcgt ctggcgact 
ggttccttc ctgaagcgg aatccgggg gttcccgag 
ggatacagg tttttcccc ccgaaacac gagactttt 
ttcaaagat gacgcaaac cacgacccc caatggtcc 
agcgagcag attatctcg gcgaagttc aaggtcaat 
ggcttggag attgggccc actgaggtt gatatcatg 
acgcggatg tcgattttg gtttttgaa attttggaa 
aaagtctgg gcgtctaga aattgcgct cttatcgat 
atgaaaata gaattcggt gtcgacaaa tcaggggaa 
atcctcgtg gccgacgtc atcgactcg gacagctgg 
cggctgtgg ccctccggg gacaagcgc ctcatggtc 
gacaagcaa gtctaccgt aacctcacc accgtcacc 
tcctccgac ctggacacc gtcaagcgc aacttcgag 
tggatcgtc gaccagctg gaccacctc gccccgccc 
aacgaccac ctggtggtc atcctcatg ggcagcccc 
tcggacgcc gaccactgc aaaaccatc cagaaacac 
tgcgaaaaa ctcggtctc aacaccgaa ctgcgcgtc 
acctcggca cacaaaggc acagaagaa acgatcaaa 
attatcaga tattacgag tctctccct ctgaaggtc 
gtttttatc gcagtggcg ggccggtcc aacggcttg 
ggaccagtg gtttcgggg aattccccg ttcccggtc 
attaactgt cccccactt aaggccgaa aatgtggac 
agggacgtg tggtccagc ttgaatgtg ccctcgggg 
ctggggtgc agtacggtc ctgtatccc gaagctgcc 
gccttaagt gcggccaat attatcggg ctgggaaat 
tatgtggtc tggagtagg ctgagagtg agacagttg 
gataattat gtcactctg gccaaggct gataagggg 
gtgaagaat aattga
    mRNA sequence
    • >TC016225 RA
      gcacgaatc atctacgat ttataaaaa caaacaacg 
gtcgctttt cgtcctctt gcacgtttt ctgcaagaa 
ttgtgtttt tgttgccat ggcgacggt caaacaggt 
cggtgacta caaattggg ggccctcat catcgaggg 
caagaccaa gcaagtgta cgagttgcc gggatcccc 
cgatttgtg ccttttgct gaataagga ccggatcac 
ggcaggcga tggcgtcaa agcgcacga tctggcggg 
aaaggcggc aatttcgaa taagacaaa tggaaaagt 
tttcgaaat tttaaattc agtcggggt gaaaacttc 
gttcgttaa agccgcctc tgatgtggc ttttatttc 
gaaaaagtg tgagatgat ccccattga gtgggtgac 
aaggcgtct ggcgactgg ttccttcct gaagcggaa 
tccgggggt tcccgaggg atacaggtt tttcccccc 
gaaacacga gactttttt caaagatga cgcaaacca 
cgaccccca atggtccag cgagcagat tatctcggc 
gaagttcaa ggtcaatgg cttggagat tgggcccac 
tgaggttga tatcatgac gcggatgtc gattttggt 
ttttgaaat tttggaaaa agtctgggc gtctagaaa 
ttgcgctct tatcgatat gaaaataga attcggtgt 
cgacaaatc aggggaaat cctcgtggc cgacgtcat 
cgactcgga cagctggcg gctgtggcc ctccgggga 
caagcgcct catggtcga caagcaagt ctaccgtaa 
cctcaccac cgtcacctc ctccgacct ggacaccgt 
caagcgcaa cttcgagtg gatcgtcga ccagctgga 
ccacctcgc cccgcccaa cgaccacct ggtggtcat 
cctcatggg cagcccctc ggacgccga ccactgcaa 
aaccatcca gaaacactg cgaaaaact cggtctcaa 
caccgaact gcgcgtcac ctcggcaca caaaggcac 
agaagaaac gatcaaaat tatcagata ttacgagtc 
tctccctct gaaggtcgt ttttatcgc agtggcggg 
ccggtccaa cggcttggg accagtggt ttcggggaa 
ttccccgtt cccggtcat taactgtcc cccacttaa 
ggccgaaaa tgtggacag ggacgtgtg gtccagctt 
gaatgtgcc ctcggggct ggggtgcag tacggtcct 
gtatcccga agctgccgc cttaagtgc ggccaatat 
tatcgggct gggaaatta tgtggtctg gagtaggct 
gagagtgag acagttgga taattatgt cactctggc 
caaggctga taagggggt gaagaataa ttgaacaaa 
tgccatttt ttatttcgt gttttattt aaaaaaata 
cataaaaag taataat
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY