| General information | ||||
| iB_03800 | iB fragment
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. |
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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.
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)adult lethality - Number of animals showing the phenotype: 2 (9 dpi)
larval musculature mostly shortened - Penetrance (percentage of animals/eggs on slide): 30% - 50%
affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 30% - 50%
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.
Lethalities 11 days after larval injection: 0.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 0.0% (includes death as larva, prepupa, pupa, adult)
- >TC004621 RA
MQSSGGNGF NALPESGTL PPMGPPAVP QNRFEPYFD AMTPRNVTA LVGKSAYLS CRVRNLGNK TVSWIRHRD IHILTVGSY TYTSDQRFQ ANHHPETDE WTLQIKWAQ KRDAGIYEC QISTQPVRS YFVNLNVVV PTAQILGGP DLHVDKGST INLTCSIRF SPEPPAYIF WYHRDDVIS YDSNRGGVS VITEKGDVT TSFLLIQHA DISDSGKYS CSPSNADVA SVRVHVLNG ERPAAMQTG SAGLSNSSL NILALMIIS YIYTAIYLS LQLETKQTV AALR
- >TC004621 RB
MQSSGGNGF NALPESGTL PPMGPPAVP QNRFEPYFD AMTPRNVTA LVGKSAYLS CRVRNLGNK TVSWIRHRD IHILTVGSY TYTSDQRFQ ANHHPETDE WTLQIKWAQ KRDAGIYEC QISTQPVRS YFVNLNVVV SEVPAGDEP SREVANKKN CSSCSPGAR LFMYAIPTA QILGGPDLH VDKGSTINL TCSIRFSPE PPAYIFWYH RDDVISYDS NRGGVSVIT EKGDVTTSF LLIQHADIS DSGKYSCSP SNADVASVR VHVLNGERP AAMQTGSAG LSNSSLNIL ALMIISYIY TAIYLSLQL ETKQTVAAL R
- >TC004621 RA
atgcaaagt tctggtggt aacggtttt aacgccctt ccagagagc ggcactctg ccgcctatg ggaccacct gcagttcca caaaatcgc tttgaacct tattttgac gcaatgact ccacgcaat gtaacagct cttgttgga aaatcggct tatctcagc tgccgcgta cgaaatttg ggaaataaa acggtttcg tggattcgt catcgggat attcacatt ttaactgtc ggcagttac acatacacg agcgaccag agatttcag gcgaatcat caccctgaa accgacgaa tggacttta cagatcaag tgggcacaa aagagagat gccggaatt tacgagtgt caaattagc acacaacca gtgaggagc tatttcgtt aaccttaat gttgtcgtt cccacagcg cagatactt gggggacct gatctgcat gtggataaa ggtagcacc atcaacttg acatgttcg atccggttc agcccggaa ccgccggct tacatcttc tggtaccac cgagacgac gtaataagt tacgattcg aacagaggc ggcgtgagc gtcatcacc gaaaaggga gatgtgact acaagtttc ttgttaatc cagcatgct gacatttca gactccggc aaatattct tgcagtccg tcgaacgca gatgttgca agcgtccga gtacacgtc ttgaacggg gagagacct gccgctatg caaaccgga tcagcggga ctgtcgaac agttctttg aatatcctc gccctaatg atcatttct tacatttac accgctatc tatttgagt ctccaactt gaaaccaaa caaacggtg gctgcctta aggtag
- >TC004621 RB
atgcaaagt tctggtggt aacggtttt aacgccctt ccagagagc ggcactctg ccgcctatg ggaccacct gcagttcca caaaatcgc tttgaacct tattttgac gcaatgact ccacgcaat gtaacagct cttgttgga aaatcggct tatctcagc tgccgcgta cgaaatttg ggaaataaa acggtttcg tggattcgt catcgggat attcacatt ttaactgtc ggcagttac acatacacg agcgaccag agatttcag gcgaatcat caccctgaa accgacgaa tggacttta cagatcaag tgggcacaa aagagagat gccggaatt tacgagtgt caaattagc acacaacca gtgaggagc tatttcgtt aaccttaat gttgtcgtt tcagaggtc ccagccggc gacgagccc tcccgtgaa gtagcgaat aaaaagaat tgtagtagt tgttcccca ggtgccagg ttgtttatg tatgccatt cccacagcg cagatactt gggggacct gatctgcat gtggataaa ggtagcacc atcaacttg acatgttcg atccggttc agcccggaa ccgccggct tacatcttc tggtaccac cgagacgac gtaataagt tacgattcg aacagaggc ggcgtgagc gtcatcacc gaaaaggga gatgtgact acaagtttc ttgttaatc cagcatgct gacatttca gactccggc aaatattct tgcagtccg tcgaacgca gatgttgca agcgtccga gtacacgtc ttgaacggg gagagacct gccgctatg caaaccgga tcagcggga ctgtcgaac agttctttg aatatcctc gccctaatg atcatttct tacatttac accgctatc tatttgagt ctccaactt gaaaccaaa caaacggtg gctgcctta aggtag
- >TC004621 RA
atttacgac actccatcc acaatttac ctgttacga acacgcgga tattttcat ttttaaata actcactcc aagtggaac aactttccg tgacaacac ccccgtccg gggcattct cccgcctct cagcggacg caacaatgt gccctcttt tattttgca aattcgtgg ataattttc gaaatcaaa cactgtgaa tccaataaa gcaggcgtg gcctaattt taactgcaa cagtgttgt cgttcgaat gcaacttgg attttagtg gcagtgaat ggaaatgtg agttttcag cgagatact ggacatttt gccgaaatg ggagacatt atggtgcca taccttctg gcggttgct atggtcttt tccaacaca gggtcgatt ctggtaaac ggagggatg caaagttct ggtggtaac ggttttaac gcccttcca gagagcggc actctgccg cctatggga ccacctgca gttccacaa aatcgcttt gaaccttat tttgacgca atgactcca cgcaatgta acagctctt gttggaaaa tcggcttat ctcagctgc cgcgtacga aatttggga aataaaacg gtttcgtgg attcgtcat cgggatatt cacatttta actgtcggc agttacaca tacacgagc gaccagaga tttcaggcg aatcatcac cctgaaacc gacgaatgg actttacag atcaagtgg gcacaaaag agagatgcc ggaatttac gagtgtcaa attagcaca caaccagtg aggagctat ttcgttaac cttaatgtt gtcgttccc acagcgcag atacttggg ggacctgat ctgcatgtg gataaaggt agcaccatc aacttgaca tgttcgatc cggttcagc ccggaaccg ccggcttac atcttctgg taccaccga gacgacgta ataagttac gattcgaac agaggcggc gtgagcgtc atcaccgaa aagggagat gtgactaca agtttcttg ttaatccag catgctgac atttcagac tccggcaaa tattcttgc agtccgtcg aacgcagat gttgcaagc gtccgagta cacgtcttg aacggggag agacctgcc gctatgcaa accggatca gcgggactg tcgaacagt tctttgaat atcctcgcc ctaatgatc atttcttac atttacacc gctatctat ttgagtctc caacttgaa accaaacaa acggtggct gccttaagg tagccattt ggcgccaat gtcaataaa tgtatataa attaagtaa aagcgcgag ttctcaacg acttataca agaaaaaat atataaaaa tacactaaa tcactgtac agaaacatt aaatttaat caagtgttt caaatttct gtgttctag agtttacct acgttgtgt gctgttatc ggatcaaat atttggatc agtaattta tgttagacg aacaaattt attttacat tgttctgtt ccgcagtaa tagtcatac tgttgaatt aacatgttg gtgtttttc gtgttgcat accacttgg agtaactta aagtgtcgg tgagaacca aaattgaaa cacttaaag ttatgaatg ttaattgtt attttaaag gttaaccat agtgtaaat ataacattt taaactgga ttaggcgat agatgtaaa attgagtaa atagaatca taacgaggg aaggttttt attggggaa gtttgtgtg aaatcatag agaaacaaa cagtaactt ccccggcaa gtgaaatat atattttat ttgttgttg acatgcaag ctataactt tgtattagg aaaatgaag gaaagtttt attggatga acaaaagtg aatagactt tcagtgaca cttttcaat gtgtttaaa aaaggtgta taattaagg atttcacgt agaaaactt cttgttgcc agaggaaag ttttgttaa ttattctcc ccaggcgaa ctggaataa attttatcc actcaacca gtatggata tttatatat taacgggtt attaatttc gcttcaaca gggtatacg taattattt cggttattt ttaaaatgt ataacattg taattattc atgttcatg tagattggc cttttgttg acggtgaaa caataaatc tcaacaaaa tggattaat cattattcg ataattatt tcccagttt gaagtgact gcagtgact ttaacgcat tttggaaga taatgtaaa ttactgtaa atactttag atagaacta acatataag ccactgctg tgtataaaa tacaggggc ggcgctaga cctttttgt attttttct atttccttg tcataagtc gtgcatcaa tgccgattt tcccaaaaa aacaaaacg ctagcgacg tcattgtta tcgaataca atgaaaaat cagtaaagc aagttacta agatagggt aaattagat ctctttgtt ttactgttc taaatgtat tcaaattac agaagtacc tatatcgac aattcatta tctcaaaat gtgtagtgt gtaatttat attactgta aagtttatt aataaataa acacccatt ttaaaa
- >TC004621 RB
atttacgac actccatcc acaatttac ctgttacga acacgcgga tattttcat ttttaaata actcactcc aagtggaac aactttccg tgacaacac ccccgtccg gggcattct cccgcctct cagcggacg caacaatgt gccctcttt tattttgca aattcgtgg ataattttc gaaatcaaa cactgtgaa tccaataaa gcaggcgtg gcctaattt taactgcaa cagtgttgt cgttcgaat gcaacttgg attttagtg gcagtgaat ggaaatgtg agttttcag cgagatact ggacatttt gccgaaatg ggagacatt atggtgcca taccttctg gcggttgct atggtcttt tccaacaca gggtcgatt ctggtaaac ggagggatg caaagttct ggtggtaac ggttttaac gcccttcca gagagcggc actctgccg cctatggga ccacctgca gttccacaa aatcgcttt gaaccttat tttgacgca atgactcca cgcaatgta acagctctt gttggaaaa tcggcttat ctcagctgc cgcgtacga aatttggga aataaaacg gtttcgtgg attcgtcat cgggatatt cacatttta actgtcggc agttacaca tacacgagc gaccagaga tttcaggcg aatcatcac cctgaaacc gacgaatgg actttacag atcaagtgg gcacaaaag agagatgcc ggaatttac gagtgtcaa attagcaca caaccagtg aggagctat ttcgttaac cttaatgtt gtcgtttca gaggtccca gccggcgac gagccctcc cgtgaagta gcgaataaa aagaattgt agtagttgt tccccaggt gccaggttg tttatgtat gccattccc acagcgcag atacttggg ggacctgat ctgcatgtg gataaaggt agcaccatc aacttgaca tgttcgatc cggttcagc ccggaaccg ccggcttac atcttctgg taccaccga gacgacgta ataagttac gattcgaac agaggcggc gtgagcgtc atcaccgaa aagggagat gtgactaca agtttcttg ttaatccag catgctgac atttcagac tccggcaaa tattcttgc agtccgtcg aacgcagat gttgcaagc gtccgagta cacgtcttg aacggggag agacctgcc gctatgcaa accggatca gcgggactg tcgaacagt tctttgaat atcctcgcc ctaatgatc atttcttac atttacacc gctatctat ttgagtctc caacttgaa accaaacaa acggtggct gccttaagg tagccattt ggcgccaat gtcaataaa tgtatataa attaagtaa aagcgcgag ttctcaacg acttataca agaaaaaat atataaaaa tacactaaa tcactgtac agaaacatt aaatttaat caagtgttt caaatttct gtgttctag agtttacct acgttgtgt gctgttatc ggatcaaat atttggatc agtaattta tgttagacg aacaaattt attttacat tgttctgtt ccgcagtaa tagtcatac tgttgaatt aacatgttg gtgtttttc gtgttgcat accacttgg agtaactta aagtgtcgg tgagaacca aaattgaaa cacttaaag ttatgaatg ttaattgtt attttaaag gttaaccat agtgtaaat ataacattt taaactgga ttaggcgat agatgtaaa attgagtaa atagaatca taacgaggg aaggttttt attggggaa gtttgtgtg aaatcatag agaaacaaa cagtaactt ccccggcaa gtgaaatat atattttat ttgttgttg acatgcaag ctataactt tgtattagg aaaatgaag gaaagtttt attggatga acaaaagtg aatagactt tcagtgaca cttttcaat gtgtttaaa aaaggtgta taattaagg atttcacgt agaaaactt cttgttgcc agaggaaag ttttgttaa ttattctcc ccaggcgaa ctggaataa attttatcc actcaacca gtatggata tttatatat taacgggtt attaatttc gcttcaaca gggtatacg taattattt cggttattt ttaaaatgt ataacattg taattattc atgttcatg tagattggc cttttgttg acggtgaaa caataaatc tcaacaaaa tggattaat cattattcg ataattatt tcccagttt gaagtgact gcagtgact ttaacgcat tttggaaga taatgtaaa ttactgtaa atactttag atagaacta acatataag ccactgctg tgtataaaa tacaggggc ggcgctaga cctttttgt attttttct atttccttg tcataagtc gtgcatcaa tgccgattt tcccaaaaa aacaaaacg ctagcgacg tcattgtta tcgaataca atgaaaaat cagtaaagc aagttacta agatagggt aaattagat ctctttgtt ttactgttc taaatgtat tcaaattac agaagtacc tatatcgac aattcatta tctcaaaat gtgtagtgt gtaatttat attactgta aagtttatt aataaataa acacccatt ttaaaa