TC009967
Fly Ortholog:
Rae1
Student Name:
Bradley Davis and Kayla Hanson
School Name:
Miami University
Year of Study:
2018
Gene Information:
Rae 1 was first identified as an RNA exporter in yeast. It is part of the Wd40-repeat B propeller protein superfamily and works as a poly A mRNA exporter in Drosophila. It is a known cell cycle regulator, and is also involved in proliferation, organization, biogenesis, and in male meiosis and male germ-cell post-meiotic differentiation in Drosophila. It is primarily located in the nucleus and endomembrane system, or as part of the nuclear pore complex (gene group membership), facilitating nucleocytoplasmic trafficking. There is experimental evidence that it works in protein binding. There is only one polypeptide and one protein coding transcript associated with this gene. (KD)
NCBI Link:
iBeetle Link:
Tribolium Transcript: 5' UTR:
TGGTTGTCGTCGTGCCTCGTTTGACAGTTGTTTGACATTTCCAACTGCGCTGTTTGTTTTTATTCGGTGAAACTTTTACAAATTATAGGAA
Tribolium Transcript: CDS:
ATGTTTAATCAGAGTTCAATCGGCGGCTTTGGGGGCACCCCCACGCCGAGTTCAGCAAACCCGATGAAGGACTTCGAAGTAACGCAGCCCCCGGACGATTCGATAAGTTCTTTGGCGTTCTCTCCGGCAACGCTCCCCCAGAATTTTTTAATAGCCGCAAGTTGGGACAACAACGTGCGTTGTTGGGAAATCGAGCAAACGGGGAAAAGCGTCCCCAAATCCATGCAAGCGTGTGGGGGCCCCGTACTTGACGTGTGTTGGAGTGACGATGGCACAAAAGTGTTTATGGCGGGGTGTGATAAACAGGCCAAGGCCTGGGATTTGGGCAGTAACCAGGTGATACAAGTTGCGCAACATGATGCCCCCATTAAGACTTGTCACTGGATAAAGGCTTCGAACTATTCGTGTTTAATGACCGGGTCTTGGGACAAGACTCTCAAATTTTGGGACACGCGCACCCCTAACCCCATGATGAGTATTAATCTACCTGAGAGGTGTTACTGTGCCGATGTGGATTACCCTATGGCGGTGGTTGGGACCGCAGGGCGCCAGATCATTGTCTACCAGCTAGACGGCAAGCCCCAGCAGTACAAAGTGCTTGATAGCCCCCTCAAATACCAGCACCGCTGTGTGGCCATTTTTAGGGATAAAAAGAAGTCTCCAACTGGGTACGCTTTAGGTTCGGTGGAAGGCCGCGTCGCCATCCAATACGTTAACCCGGCAAACCCCAAAGACAATTTTACGTTTAAGTGCCACAGATCCAACGGGGCACCGAACGGCTATCAGGACATTTACGCCGTCAATGACATTGCATTCCACCCAGTCCATGGCACTTTGGCCACTGTTGGGTCCGATGGGTCTTTCAGTTTCTGGGATAAGGACGCACGCACCAAATTGAAATCTTCTGACCTTATGGAACAACCAATCACGCGGTGCGCTTTCAACAGCAATGGGCAAATCTTCGCCTACGCTGTTAGCTACGACTGGTCCAAAGGACACGAATTTTACAATTCAACCAAGAAGAATTACATTTTCTTGCGGTCATGTTATGATGAATTGAAACCACGAACCTCTTCATAA
Tribolium Transcript: 3' UTR:
TTGCTTACTTTTTTCAACTGTAATCAGGCTTAATAATTTGGTCCACATTATGTATTTACATTTATTTAAACAATTACAAGTTACTACAAGCA
Tribolium Protein:
MFNQSSIGGFGGTPTPSSANPMKDFEVTQPPDDSISSLAFSPATLPQNFLIAASWDNNVRCWEIEQTGKSVPKSMQACGGPVLDVCWSDDGTKVFMAGCDKQAKAWDLGSNQVIQVAQHDAPIKTCHWIKASNYSCLMTGSWDKTLKFWDTRTPNPMMSINLPERCYCADVDYPMAVVGTAGRQIIVYQLDGKPQQYKVLDSPLKYQHRCVAIFRDKKKSPTGYALGSVEGRVAIQYVNPANPKDNFTFKCHRSNGAPNGYQDIYAVNDIAFHPVHGTLATVGSDGSFSFWDKDARTKLKSSDLMEQPITRCAFNSNGQIFAYAVSYDWSKGHEFYNSTKKNYIFLRSCYDELKPRTSS (KD)
Forward Primer 1:
GAGTTCAATCGGCGGCTTTG
Reverse Primer 1:
TGGCCTGTTTATCACACCCC
Forward Primer 2:
GTGCCGATGTGGATTACCCT
Reverse Primer 2:
GCGAAGATTTGCCCATTGCT
Gel Picture:
Cloned Fragment:
GTGCCGATGTGGATTACCCTATGGCGGTGGTTGGGACCGCAGGGCGCCAGATCATTGTCTACCAGCTAGACGGCAAGCCCCAGCAGTACAAAGTGCTTGATAGCCCCCTCAAATACCAGCACCGCTGTGTGGCCATTTTTAGGGATAAAAAGAAGTCTCCAACTGGGTACGCTTTAGGTTCGGTGGAAGGCCGCGTCGCCATCCAATACGTTAACCCGGCAAACCCCAAAGACAATTTTACGTTTAAGTGCCACAGATCCAACGGGGCACCGAACGGCTATCAGGACATTTACGCCGTCAATGACATTGCATTCCACCCAGTCCATGGCACTTTGGCCACTGTTGGGTCCGATGGGTCTTTCAGTTTCTGGGATAAGGACGCACGCACCAAATTGAAATCTTCTGACCTTATGGAACAACCAATCACGCGGTGCGCTTTCAACAGCAATGGGCAAATCTTCGC (KD)
Injection concentration:
1ug/ul
Number Injected:
21
Number Survived to Adult:
0
Number w/ RNAi Phenotype:
0
Percent Penetrance:
0
RNAi Phenotype Description:
No obvious phenotype directly due to knockdown of Rae1, completely lethal by end of pupal stage.
RNAi Phenotype/Assay Images:
Discussion:
None of the beetles survived past late pupal stage. Some of the beetles had strange patterns of wing translucence, but this is likely due to problems with the development of the pupa caused by a cascade of effects from Rae1 knockdown. In Drosophila, knockdown of Rae1 shows a decrease in organ size while an up-regulation of Rae1 causes an increase in organ size. The gene is very highly conserved between the two species, so it is likely that Rae1 is very important in the life cycle of both species. It is clear that the gene is essential for survival in Tribolium, further explaining its evolutionary importance.
Notebook:
Annotated Genbank File:
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