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Gene information
General information
TC003902 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_007418.3: 26404525..26409209
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
    		TC003902
    iBeetle screen iB_00633

    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: 0.0% (includes death as pupa, adult)

    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): 30% - 50%
    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): 50% - 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): 50% - 80%

    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: 0.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 30.0% (includes death as larva, prepupa, pupa, adult)

    adult lethality - Number of animals showing the phenotype: 2 (19 dpi)
    Days post injection

    adult lethality - Number of animals showing the phenotype: 1 (22 dpi)
    Days post injection

    Adult morphology
    elytra not closed - Number of animals showing the phenotype: 4


    elytra partially blistered - Number of animals showing the phenotype: 4

    Fertility
    egg production: number <50% of expected -
    mature egg deposition blocked number 1st lateral oviduct: >5 number 2nd lateral oviduct: - Number of animals showing the phenotype: 4
    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC003902 RA
      MSVPNDQEN YSKAVEKNP KSGNRAALS NLENILPQN 
AFNIRIKKT IKTTATETK IRSASASEA TTQVKFAYK 
KENITERKN GEFNIDEVF VEGEYHYLT YLKPEENRP 
KTKSEDLGE KIDRTLPMV RVTAPTCER KLEVKKPAK 
KQTLRKFSN RRKQANLEE LFRNCLYEE DYFDVNFRH 
MLLEEGNYV YRKQFLKKS IDENYRMLV VNWLVYAHQ 
VLELSENTF FVAVRMFDY VVASQKTAG NKYILTAIT 
STWIANKMF DVRQIHISK LFALCNGKF SNQQFLSTE 
REILKVFKF DINISEPSS FLFYFVKLE NLELDSKIC 
YSALFLLEI FALLLDFSS LPTSFLAAV ALYLVLVKY 
ELDTKLHLE VAETYCLDK ESFVRKSEQ LALCVEHLL 
KEKNASSEI VAKYSTEMR GQVARNFVS
    CDS sequence
    • >TC003902 RA
      atgtctgtt ccaaatgac caagaaaat tactcaaaa 
gccgtcgaa aaaaatccc aaatcggga aaccgtgcc 
gccctctcc aacctcgaa aacatcctc cctcagaac 
gcatttaac atccgcatt aagaaaaca attaagaca 
actgcaacc gaaacgaaa attcgctca gcttcggcc 
agtgaagcc accacccag gtgaagttc gcctacaag 
aaagaaaac atcaccgag agaaaaaac ggggagttt 
aacatcgac gaagtcttc gtcgagggg gagtaccac 
tacctcact tacctcaag cctgaggaa aataggcca 
aagacaaag tctgaagat ttgggcgag aaaattgat 
cgaacttta ccgatggtg agagttacg gcaccgact 
tgtgagagg aagctcgag gtgaagaaa ccagccaag 
aagcagacg ctcaggaaa ttcagcaac aggcgcaaa 
caggccaat ttggaagaa ctgtttcgg aactgtttg 
tacgaggag gactatttt gatgtgaac ttccggcat 
atgctttta gaagagggc aactacgtc taccgcaaa 
caatttttg aaaaaatcc atcgacgag aattacaga 
atgctcgtc gtcaattgg ctggtctac gcccatcaa 
gtgttggaa ttgtccgaa aacacgttc ttcgttgct 
gtccgaatg ttcgattat gttgtagca tctcagaaa 
acggctgga aataaatac attttaacc gctatcacg 
agcacttgg atcgctaac aaaatgttc gatgttcgt 
caaatacat atctccaaa ctgttcgcc ttgtgcaac 
ggcaagttt tccaatcag cagtttcta tcaacggag 
cgggaaatc ttgaaagtg ttcaaattc gatattaac 
atctctgaa cccagttcg tttctgttt tatttcgtc 
aagttggag aatttagag ctggactca aaaatttgc 
tacagtgcg ttgtttctc ctcgaaatt tttgcttta 
cttttggac ttttcctct ctgccgacg tcgtttctc 
gctgctgtg gccttgtat ctagttttg gtaaaatac 
gagcttgac accaaattg catctggaa gtggcggaa 
acatactgt cttgacaaa gaatcgttt gtacggaag 
tccgagcaa cttgcactt tgtgttgag catttgttg 
aaggaaaag aatgcgagc agcgaaatt gtggccaaa 
tattcgacc gaaatgagg ggtcaagtc gctcgtaat 
tttgtgtct taa
    mRNA sequence
    • >TC003902 RA
      ttcttcgac taaagatac gatctagaa agaacattt 
ttgccagtg gaccacaag cagtgtaac aaagtgcaa 
aatgtctgt tccaaatga ccaagaaaa ttactcaaa 
agccgtcga aaaaaatcc caaatcggg aaaccgtgc 
cgccctctc caacctcga aaacatcct ccctcagaa 
cgcatttaa catccgcat taagaaaac aattaagac 
aactgcaac cgaaacgaa aattcgctc agcttcggc 
cagtgaagc caccaccca ggtgaagtt cgcctacaa 
gaaagaaaa catcaccga gagaaaaaa cggggagtt 
taacatcga cgaagtctt cgtcgaggg ggagtacca 
ctacctcac ttacctcaa gcctgagga aaataggcc 
aaagacaaa gtctgaaga tttgggcga gaaaattga 
tcgaacttt accgatggt gagagttac ggcaccgac 
ttgtgagag gaagctcga ggtgaagaa accagccaa 
gaagcagac gctcaggaa attcagcaa caggcgcaa 
acaggccaa tttggaaga actgtttcg gaactgttt 
gtacgagga ggactattt tgatgtgaa cttccggca 
tatgctttt agaagaggg caactacgt ctaccgcaa 
acaattttt gaaaaaatc catcgacga gaattacag 
aatgctcgt cgtcaattg gctggtcta cgcccatca 
agtgttgga attgtccga aaacacgtt cttcgttgc 
tgtccgaat gttcgatta tgttgtagc atctcagaa 
aacggctgg aaataaata cattttaac cgctatcac 
gagcacttg gatcgctaa caaaatgtt cgatgttcg 
tcaaataca tatctccaa actgttcgc cttgtgcaa 
cggcaagtt ttccaatca gcagtttct atcaacgga 
gcgggaaat cttgaaagt gttcaaatt cgatattaa 
catctctga acccagttc gtttctgtt ttatttcgt 
caagttgga gaatttaga gctggactc aaaaatttg 
ctacagtgc gttgtttct cctcgaaat ttttgcttt 
acttttgga cttttcctc tctgccgac gtcgtttct 
cgctgctgt ggccttgta tctagtttt ggtaaaata 
cgagcttga caccaaatt gcatctgga agtggcgga 
aacatactg tcttgacaa agaatcgtt tgtacggaa 
gtccgagca acttgcact ttgtgttga gcatttgtt 
gaaggaaaa gaatgcgag cagcgaaat tgtggccaa 
atattcgac cgaaatgag gggtcaagt cgctcgtaa 
ttttgtgtc ttaataatt gtttgtttt aattttaga 
ccgtgctta attattgtg atgttttac ccaatgaat 
tttacataa aatgctgga tcagttcat acttgcctt 
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