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Gene information
General information
TC009977 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_007422.5: 10795479..10803811
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
    		TC009977
    iBeetle screen iB_01638

    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: 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 with embryonic tissue, no cuticle, no fluorescence - Penetrance (percentage of animals/eggs on slide): more than 80%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): more than 80%
    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%

    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: 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

    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 2 (16 dpi)
    Days post injection

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC009977 RA
      MATNEENQN STNKKTMEQ CYENYILVD VGANLTNKK 
YSRDLESVI QRARDSGVQ KIMVTGTSG KSSKEALRL 
TRIYPGTLY STAGIHPHD AKSYTDESW NELKLVASN 
PECVAVGEC GLDYNRNFS EPSEQRQVF RKHIELAIE 
INKPLFVHE RDAHEDLLE ILDQYKGQL PPVLVHCFT 
GTAEQALTY LSRGFYIGL TGYLCKDKS DVGVRKLLV 
DGSIPLDRL LVETDAPFM YPNTRASKL PLHVKDGLT 
ERSLTFLHR YCTFQRNEP CSLPAIVEM IAAFMNKKP 
EDVALATSF NALKLFGLS Q
    CDS sequence
    • >TC009977 RA
      atggcaact aacgaagaa aatcaaaat tcgacaaac 
aagaaaacc atggaacaa tgttacgaa aattacatt 
ctggtggat gttggcgca aatttaacc aataaaaag 
tacagcagg gacctggaa tcggtcatt caaagggca 
cgggactca ggtgtccaa aaaatcatg gtcacaggc 
acctctggc aagtcaagc aaagaagcg ctaagattg 
accagaatc tatcctggc actttatat tcgactgca 
ggaatacac ccccatgac gcaaaatca tacactgac 
gaatcttgg aacgaactg aaactagtt gcctcaaac 
cccgaatgt gtggccgtg ggcgagtgc ggcctggac 
tacaatcga aatttctca gagccaagt gaacagaga 
caagttttt cgaaaacat atcgaatta gcgattgaa 
attaataaa ccgttattt gtgcacgaa cgagacgct 
catgaagat ttattagag attttggat cagtacaaa 
ggtcaatta cctcccgtt ctggtgcac tgtttcaca 
gggacggcc gaacaagcg ctgacatat ctttcaaga 
ggcttttac ataggatta acaggctac ttgtgtaaa 
gataaatct gatgttggc gttcgtaaa ttattggta 
gatgggtct ataccattg gacagactc ctagtggag 
acagatgcc ccgtttatg tacccaaac acacgagct 
tcaaaacta cctcttcat gtcaaagat ggtctaacc 
gaaagatca ttaaccttc cttcatcga tattgtacc 
ttccaaagg aacgaaccc tgctctttg ccagcgatc 
gtcgagatg attgcggct tttatgaat aagaaaccg 
gaggacgtg gccttagct acgtctttt aatgccttg 
aaacttttc ggattgtct caatga
    mRNA sequence
    • >TC009977 RA
      caagagaaa atcactaca ttaatatca acagctgtt 
tacttttat ttctgtcac aatgacaca tgactaatg 
gggttatat ttgtttctg cagtcattt tgaaccact 
ttattatca tttcgtgta aaattaaac cgttccaag 
aacatggca actaacgaa gaaaatcaa aattcgaca 
aacaagaaa accatggaa caatgttac gaaaattac 
attctggtg gatgttggc gcaaattta accaataaa 
aagtacagc agggacctg gaatcggtc attcaaagg 
gcacgggac tcaggtgtc caaaaaatc atggtcaca 
ggcacctct ggcaagtca agcaaagaa gcgctaaga 
ttgaccaga atctatcct ggcacttta tattcgact 
gcaggaata cacccccat gacgcaaaa tcatacact 
gacgaatct tggaacgaa ctgaaacta gttgcctca 
aaccccgaa tgtgtggcc gtgggcgag tgcggcctg 
gactacaat cgaaatttc tcagagcca agtgaacag 
agacaagtt tttcgaaaa catatcgaa ttagcgatt 
gaaattaat aaaccgtta tttgtgcac gaacgagac 
gctcatgaa gatttatta gagattttg gatcagtac 
aaaggtcaa ttacctccc gttctggtg cactgtttc 
acagggacg gccgaacaa gcgctgaca tatctttca 
agaggcttt tacatagga ttaacaggc tacttgtgt 
aaagataaa tctgatgtt ggcgttcgt aaattattg 
gtagatggg tctatacca ttggacaga ctcctagtg 
gagacagat gccccgttt atgtaccca aacacacga 
gcttcaaaa ctacctctt catgtcaaa gatggtcta 
accgaaaga tcattaacc ttccttcat cgatattgt 
accttccaa aggaacgaa ccctgctct ttgccagcg 
atcgtcgag atgattgcg gcttttatg aataagaaa 
ccggaggac gtggcctta gctacgtct tttaatgcc 
ttgaaactt ttcggattg tctcaatga tccgaaagt 
tgtactaat taaggggaa catacaaac atattatat 
tttgtctat tttcgaaat catgtgggc agctttatt 
aaacgaatt tagctataa tagggcgaa gaatacaac 
gaatcgtgt aaataattt taggtgatc accaatgtc 
aacatgttt accaaaagc ggttattac cgttttgga 
actataaag tttatcacg tcttatcct gtatttttg 
tagttggac tacgttacg tttagcaac tgttaaagc 
cgaattgta tattttagg taacatgaa atgtatttg 
ttgactgct tagttttat aatgcggac gaaagattt 
atgttgaag taatagaat tacgctttg ttaaagtat 
cataatttt gaattgttg atataggaa gtatgaaat 
tatggcgta taatcaatc tgatgcaat atttttgtt 
gttgctgta atttcggta gcatgttct tgtagtttt 
aattttatg gacgctgtg tagtgtaaa atggcccat 
tatttttgt aattaattt tcatatttt aggggtttg 
tttgccaca ataattgcc tctttatgt gatatttgt 
ttccatttt atagacaca aatgtgcct tatctatta 
caatatgaa gaataaata agttgtaat gaa
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY