TC003570
Fly Ortholog:
Shaven
Student Name:
Megan Lytle and Micah Morris
School Name:
Miami University
Year of Study:
2017
Gene Information:
Name: shaven; transcription factor involved in bristle development
iBeetle Link:
Tribolium Transcript: 5' UTR:
cataacagtgcgacgggcactcactcgtccggcacagtcggaacagagaagtcagtggatcgcatccttcccgaaagcccagtcggcgtggtcctcgttccggaatcttatcacaaactgagaaaaatcctccggaaccgacacagttcatcgcctcacccggcatcgatatcttccggttagtgttctgtgctggcgttgatatctccgggattttcagtgcttccgccgggtctagtgccgatttcgatgcgttcaggacgagtactctatcccagtgacacaatcgactatttttaaggataattgaacgagtgtagtgcgctaattaatcttctgtgatcgatgaaatgttactatgacgtgcttcaaccgcacaaaaaactaacatgttataatattaatggtcgcgattgacttggaccgagatttgtgattggtgcgtgtgaattgttacgttcgagtgattggttaaggccctttttcatttagatttttaagtgcggtcccatcattgagggcaat
Tribolium Transcript: CDS:
atgatcacgatggagtgccagccagggggtcatggagcacacctggggcactccatgatggtggaccccacgtcggggtcgatgcatcatcaggagcaggagcacaagaaaaagcgagacggaggctcacatgttggatcttcttcaggagacggtcatgggggcgtcaatcagctgggcggagtgttcgttaacggtaggccgctgccggacgttgtcaggcagaggatagtcgaattggcccacaacggagtacggccctgtgatatttcaaggcagctgagggtgtcccacggctgcgtctccaaaatcttatcaaggacaaatgtttgttgttccagatattacgaaacgggtagttttaaggcaggtgtgatcggcggctccaaacccaaagtagcaacaccccctgtagtagacgccatagccaactacaaaagagaaaacccaacgatgttcgcttgggaaatccgtgatagacttttagctgaaggcatttgctctcaagataacgtacctagtgtttcatcgataaataggattgtaagaaataaagcggccgaaaaagcgaaacatgcacaccaaatagcgaccagcggaggaacgacgtcttcgtcagggggtcccgtctccgttatcgcacacgcacctgcaacgcaacttacggaacctcgctcaggagcttatagtataaacgggatcctcggcctccagcacgaccccaacgggaacagcatcaagaggaaaaggatagaggatcacgatgaaaatagagatataaacggacatccggaggatgacctgaaaagacagcgatcgcagtataatggagaccagctttattcaaatctgtggtcgagcaagtggagcatcaaagacgagcacaaactcctctcggagctgggaggcgcgggcacaacgacgggccagacgggatactacgacacccatggcggcttccccagcgtgggggtggcgagctcggcggcggacatctacgacaccatcaacaccatgagccaagccacctcgcagaacctctacacgccgcccctggcgggctctctggggaccagttcattaactccattagcacctatcactatgcatgagatgaaactagaatccactaggataatggatcccgcaatgtccccttaccattcagccgagactagctcaccgtacagtggcgtggtcggtctgggggcagtggtagacggggcttcccccggtttaccactagccttaccctccgaaccaggaagtcagcccacgtcccatacagggtcgcccgatcccgccgcattgactgtgctccagccatccaatggctccaaccagcattactcgaccatgttgccaagctttggctattccacaggtaccactgtggtgccagggactgattacgcgtacagtacggcttatagtcagtacggcaccgcttacggcacctacggctacggaagtgcctccagcgggttactcagtaagtaa
Tribolium Transcript: 3' UTR:
acaattcgccgtactactatacaaacaacgacgcccctgtgaacaaccacctgaactgcacaagctcgactgaaaacgctgaacagaatagcc
Tribolium Protein:
MITMECQPGGHGAHLGHSMMVDPTSGSMHHQEQEHKKKRDGGSHVGSSSGDGHGGVNQLGGVFVNGRPLPDVVRQRIVELAHNGVRPCDISRQLRVSHGCVSKILSRTNVCCSRYYETGSFKAGVIGGSKPKVATPPVVDAIANYKRENPTMFAWEIRDRLLAEGICSQDNVPSVSSINRIVRNKAAEKAKHAHQIATSGGTTSSSGGPVSVIAHAPATQLTEPRSGAYSINGILGLQHDPNGNSIKRKRIEDHDENRDINGHPEDDLKRQRSQYNGDQLYSNLWSSKWSIKDEHKLLSELGGAGTTTGQTGYYDTHGGFPSVGVASSAADIYDTINTMSQATSQNLYTPPLAGSLGTSSLTPLAPITMHEMKLESTRIMDPAMSPYHSAETSSPYSGVVGLGAVVDGASPGLPLALPSEPGSQPTSHTGSPDPAALTVLQPSNGSNQHYSTMLPSFGYSTGTTVVPGTDYAYSTAYSQYGTAYGTYGYGSASSGLLSK
Forward Primer 1:
GCGGACATCTACGACACCAT
Reverse Primer 1:
CTCGGCTGAATGGTAAGGGG
Forward Primer 2:
CTTCAGGAGACGGTCATGGG
Reverse Primer 2:
GGGGTGTTGCTACTTTGGGT
Gel Picture:
Cloned Fragment:
CTTCAGGAGACGGTCATGGGGGCGTCAATCAGCTGGGCGGAGTGTTCGTTAACGGTAGGCCGCTGCCGGACGTTGTCAGGCAGAGGATAGTCGAATTGGCCCACAACGGAGTACGGCCCTGTGATATTTCAAGGCAGCTGAGGGTGTCCCACGGCTGCGTCTCCAAAATCTTATCAAGGACAAATGTTTGTTGTTCCAGATATTACGAAACGGGTAGTTTTAAGGCAGGTGTGATCGGCGGCTCCAAACCCAAAGTAGCAACACCCC
Injection concentration:
2.6 mg/µL
Number Injected:
15
Number Survived to Adult:
0
Number w/ RNAi Phenotype:
14/15 pupa, no adults completely survived although they still appeared to have the phenotype
Percent Penetrance:
N/A
Assay Description:
Visual observation of phenotypes
RNAi Phenotype Description:
Larvae:
Loss of sensory bristles on one injected larva?
Pupae:
Urogomphi shortened/nearly missing in pupae
Gin-traps are disrupted, some nearly missing
Missing sensory bristles
Dead adult Tribolium:
It appears that most of the beetles died during adult eclosion
Pigmentation differences on abdominal sclerites; mottled
No sensory bristles
Small eyes (about half the normal size), eye structure almost completely lacking ommatidia
RNAi Phenotype/Assay Images:
Discussion:
We speculate that since the Tribolium have no sensory hairs, the eclosion process is stunted due to the fact that the beetles cannot sense their surroundings. The shaven gene appears to be involved in even more sensory processes/features than we previously expected, including sensory bristles, eyes, urogomphi, and gin traps. Based on the loss of sensory bristles and ommatidia in the eyes, and what we already know about shaven in Drosophila, it appears that the function of this gene has been conserved between the two in this regard. However, since urogomphi and gin traps are not present in Drosophila, it appears that the genes that form sensory structures may have been co-opted to help form these additional structures. Future work could look more into the role of shaven in urogompi and gin traps since this was something that was unexpected. Also, it would be interesting to try experimenting with RNAi at various concentrations.
Notebook:
Annotated Genbank File:
Powered by Drupal
