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Gene information
General information
TC006665 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_007423.3: 12690578..12693522
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
    		TC006665
    iBeetle screen iB_04189

    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: 30.0% (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: 2 (9 dpi)
    Days post injection

    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature fluorescence decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 50
    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
    • >TC006665 RA
      MEVGEVSEN PEDKIGQLE AEKKRIQDE FNMQRAKMK 
ELFLQKEDE LLRRNHENA RLNSEIEEL KRELDDLKS 
QLVVAGMTL ENSFEQEKR KANEEIATL QQLVHETVE 
ESSSSRSLY DSELKNLQT YIQQLQNEI LVLKQQKQQ 
SPHHSSQES SLAPSVVLN ALTKGIAKK LGADAFAAS 
QDSGDDPAR KVETSDHAQ EDPEVLRSL IGPLEDQIQ 
ALKEKLRAT DEQLQRCRE CGHSQDKND QNVSTNTSF 
DSPKHATSC DMCSNYEDQ LVAEQRKTK ELELKVQKA 
EKAAERHKE DLLKEIGFR KEIEEKWNE KKEEHKQQV 
AELTRSTEC AEQDLKELR QVFSQTCAE MSEMLKKLT 
KGREKVQEE LTNLQKEND NLVGKYTIH SQELQSEAI 
NLPNTVEEL HELVLKHHQ DLIIAKIGK EAAEEKVNT 
LQSDILLLR DQITNDQHE KEVIENNLA QEIDLLKKQ 
KHQLEKEKK LYLANQEKL QNTDKANLA QIAELQKQI 
HELQTIKRQ LEDQNSELR TRVSSLQQE LDTSETVQK 
DFVRLSQSL QGTYPLSSF SHPESRCNW RKSASPTPK 
SDGSTKKTS SSAPTVAPT SAAPNARST VDTAARSSA 
PRVSPELWK VGLACSRQR FVMSVILY
    CDS sequence
    • >TC006665 RA
      atggaagtg ggtgaagtc tccgaaaat ccggaagac 
aaaatcggg caattggag gccgaaaag aagcgaatc 
caggatgag ttcaacatg caaagggcc aaaatgaaa 
gaactgttc ttgcaaaaa gaagatgaa ctgttacgc 
agaaatcac gaaaacgca cgcttaaat tccgaaatc 
gaagagtta aagcgcgaa ttggacgat ttgaaaagt 
cagttagtt gtggcaggg atgaccctt gagaatagt 
tttgagcag gagaaacgt aaagccaac gaggaaatt 
gcaacgtta caacaacta gttcacgaa accgtggaa 
gagtccagc tctagtcgg agtttgtac gacagcgag 
cttaaaaac ttacaaacg tatatccag caattacaa 
aacgagatt ttagtcctg aagcagcaa aagcaacag 
tcgccccac cattccagc caggagtct agtctggct 
cctagtgtc gtccttaac gccctaact aaaggaata 
gctaaaaaa ttgggagct gacgcgttc gctgcctct 
caagacagt ggcgatgac cccgccaga aaggtagag 
accagcgac catgcccag gaggacccc gaggtgctg 
cgctcgctc attggcccc cttgaggac cagatccag 
gccctcaag gaaaagctg cgcgcgact gacgaacag 
ctacaaagg tgtcgcgag tgtggccac agccaggac 
aaaaacgac cagaatgtt tcgacaaac acgtcgttc 
gattcgccg aaacatgcc acttcgtgc gacatgtgc 
agcaattac gaggaccag ttggtggcc gaacagcgc 
aaaacgaaa gagttggaa ctgaaagta caaaaggcg 
gaaaaggcg gccgaaaga cacaaggag gatttgctc 
aaggagatc ggcttccgg aaggagatc gaggagaag 
tggaacgag aaaaaggag gaacataag cagcaagtg 
gccgaactg acccggagc acggaatgt gccgagcag 
gacttgaag gagttgcgg caggttttt agccagaca 
tgcgccgaa atgagcgaa atgttgaag aaattgacc 
aaagggcgg gagaaagtc caggaggaa ttaaccaat 
ctccaaaag gagaacgat aatttggtc gggaagtac 
acaattcac tcacaagag ctgcagagt gaagcgatt 
aatctcccg aatactgtt gaggagctt cacgagttg 
gtgctaaag caccaccag gatttgatt atagccaag 
atcgggaag gaggcggct gaggaaaaa gtcaatact 
ttgcagtcg gatatttta ttgttgcga gatcagatt 
accaacgac caacacgag aaggaagtt attgagaat 
aatctggca caagaaatt gatttgttg aagaagcag 
aaacatcag ttagagaag gagaaaaag ttgtatttg 
gccaatcag gagaagttg caaaacacg gataaggcc 
aatttggca caaattgca gagctgcag aagcaaata 
cacgagctg caaacaatc aaacgccag ctggaagac 
cagaacagc gagctccgc acgcgcgtc tcctccctg 
cagcaggag ctcgacacg agcgaaacg gtccagaaa 
gacttcgtg cgtttatcg caaagcttg cagggtacg 
taccccctg tccagtttt tctcaccct gagtccaga 
tgcaattgg agaaaatcc gcgagtccg acacccaag 
tccgatggc agcacgaag aagacgtcg agcagtgcc 
ccaactgtc gctccgact tcggcggcc ccaaacgca 
aggagcact gtcgacact gcggccaga tcttctgca 
cctcgtgtc tcaccagaa ctgtggaaa gtggggcta 
gcatgctcc cgtcaaagg tttgtgatg tctgtcata 
ctttactag
    mRNA sequence
    • >TC006665 RA
      aacaacgac aaaaacacc acaaaattc aaaaaaaca 
caccgattg cgagtaatt tgtcttcac acattaatc 
agttattac taattaaac aattgatta acgaattag 
cgaattaag cgctacata atcaattga tccgctcgg 
tcggtaaca cgtcaattg tgacaagta tcataggtt 
aggagctta atttgtgat ttaattgca aaaaccaca 
aaattatca attcgcccg taaatatta acgaacgtg 
agtcagtgc tgacgtaat gattgttta cttgtgcta 
aaccaccaa aatggaagt gggtgaagt ctccgaaaa 
tccggaaga caaaatcgg gcaattgga ggccgaaaa 
gaagcgaat ccaggatga gttcaacat gcaaagggc 
caaaatgaa agaactgtt cttgcaaaa agaagatga 
actgttacg cagaaatca cgaaaacgc acgcttaaa 
ttccgaaat cgaagagtt aaagcgcga attggacga 
tttgaaaag tcagttagt tgtggcagg gatgaccct 
tgagaatag ttttgagca ggagaaacg taaagccaa 
cgaggaaat tgcaacgtt acaacaact agttcacga 
aaccgtgga agagtccag ctctagtcg gagtttgta 
cgacagcga gcttaaaaa cttacaaac gtatatcca 
gcaattaca aaacgagat tttagtcct gaagcagca 
aaagcaaca gtcgcccca ccattccag ccaggagtc 
tagtctggc tcctagtgt cgtccttaa cgccctaac 
taaaggaat agctaaaaa attgggagc tgacgcgtt 
cgctgcctc tcaagacag tggcgatga ccccgccag 
aaaggtaga gaccagcga ccatgccca ggaggaccc 
cgaggtgct gcgctcgct cattggccc ccttgagga 
ccagatcca ggccctcaa ggaaaagct gcgcgcgac 
tgacgaaca gctacaaag gtgtcgcga gtgtggcca 
cagccagga caaaaacga ccagaatgt ttcgacaaa 
cacgtcgtt cgattcgcc gaaacatgc cacttcgtg 
cgacatgtg cagcaatta cgaggacca gttggtggc 
cgaacagcg caaaacgaa agagttgga actgaaagt 
acaaaaggc ggaaaaggc ggccgaaag acacaagga 
ggatttgct caaggagat cggcttccg gaaggagat 
cgaggagaa gtggaacga gaaaaagga ggaacataa 
gcagcaagt ggccgaact gacccggag cacggaatg 
tgccgagca ggacttgaa ggagttgcg gcaggtttt 
tagccagac atgcgccga aatgagcga aatgttgaa 
gaaattgac caaagggcg ggagaaagt ccaggagga 
attaaccaa tctccaaaa ggagaacga taatttggt 
cgggaagta cacaattca ctcacaaga gctgcagag 
tgaagcgat taatctccc gaatactgt tgaggagct 
tcacgagtt ggtgctaaa gcaccacca ggatttgat 
tatagccaa gatcgggaa ggaggcggc tgaggaaaa 
agtcaatac tttgcagtc ggatatttt attgttgcg 
agatcagat taccaacga ccaacacga gaaggaagt 
tattgagaa taatctggc acaagaaat tgatttgtt 
gaagaagca gaaacatca gttagagaa ggagaaaaa 
gttgtattt ggccaatca ggagaagtt gcaaaacac 
ggataaggc caatttggc acaaattgc agagctgca 
gaagcaaat acacgagct gcaaacaat caaacgcca 
gctggaaga ccagaacag cgagctccg cacgcgcgt 
ctcctccct gcagcagga gctcgacac gagcgaaac 
ggtccagaa agacttcgt gcgtttatc gcaaagctt 
gcagggtac gtaccccct gtccagttt ttctcaccc 
tgagtccag atgcaattg gagaaaatc cgcgagtcc 
gacacccaa gtccgatgg cagcacgaa gaagacgtc 
gagcagtgc cccaactgt cgctccgac ttcggcggc 
cccaaacgc aaggagcac tgtcgacac tgcggccag 
atcttctgc acctcgtgt ctcaccaga actgtggaa 
agtggggct agcatgctc ccgtcaaag gtttgtgat 
gtctgtcat actttacta gtcaaatcg tcggcgccc 
tacttcagc gaagcgccc ccgatgact taagcagtt 
aaggttgta ttcaaatgt ctaataaat tctgttcca 
attt
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