TC010832
Fly Ortholog:
Big brain (bib)
Student Name:
Catherine Smith and Lan Dao
School Name:
Miami University
Year of Study:
2018
Gene Information:
Big brain (bib) coded for a protein which is thought to be similar to the water- and ion-conducting aquaporin (AQP) channels. The coding protein serves as membrane channels that selectively transport water, small neutral molecules, and ions in and out of and between cells. This is base on the family this protein is in (MIP). The previous research of Drosophila pointed out loss of phenotypes of Big Brain was bigger brain size and the loss of formation of the sensory system.
NCBI Link:
iBeetle Link:
Tribolium Transcript: 5' UTR:
TCTGCAGTCCGCTGTCCGGACCGAAAAATGGTGTCCCGCGTGGGGAAGGGATGGCGGATCCGCTTGTAATTGGTGATGATCCTGGGAGGGGGAATTCACCATGTGGTCGGTCGAATGCAATGTGCGAAGAGAGGTTTCTTCGGAAATGTCCCGTCGGTCCATCGGCACCGTTCTTTAAATTAAACCTCGTCCCTTTTTCCAAAAATGAACACTTAGTGCAATATTTCGAGT
Tribolium Transcript: CDS:
ATGGCAGAGGAACGCGTTAATTTATCTAGTGAGCCGAGCATAGACTACCACATAGTGACACTTTTCGAAAAGTTGGAAGCTTTAAGAAATGATAAGCCGAACTCGCAGTTTACTTCAAAATTGTCAATTAAAGTCGAGGTCCGCACTTTGGAATTTTGGAGATCGGTCATCTCCGAGTGCATAGCGTCTTTTGTTTACGTGTTTGTGGTGTGCGGTGCAAAGGCAGGTGCTTTTGACGGAGGTTCGGCAACTACCTCTTTGGGAGTTCTTTCAACAGCACTGGCAGCTGGCTTTGCAATGATATCGTTAACACAGTGCTTCGGACACATCTCTGGGGCGCACATAAACCCGGCCGTGAGTCTGGCCATGGGCATTACGAAGAATATTTCGCTTCTGCGGACGACAATGTTCATTGTCGCACAATGCGGGGGTGCGATCGCCGGCGCCGCTTTCCTCTACGGCGTATCCATTCCGCGAACTCAGGAGGATCTCCCGATGTCCCTGCCGATAAAAGATGGGCCTTGGGAGAGGTTCGGGAACGAGTTTATTCTGACTTTCGTGGTGGTTTTGGCCTATTTCGTCTCAATGGACGCCCAAAGGAGGTGGATGGGCATGTCGGCGATTACGATCGGAGCCACGTACTCCGCTTGCTCGTTCGTTTCGCCGATGCCGCACTTGAACCCAGCGCGTTCGCTGGGGCCGTCCTTTGTTTTGAAAAAATGGGACAACCACTGGGTTTATTGGCTGGGGCCCATAATTGGCGGCGTGGCAGCTGGGTTAATCCACGAGTACATTTTCAACTCGAAGCGCTCGAAACGCCAAAACAGCTCGAGTGACGGCGACTCGTCCAGCATCCAGTCGGATGAGGACACTTACGACGACCTCGACAAACCCAATGCCCCAAAATACCAGGAATCGACTTACAACTCCTACAGAGCCGGCGCTCCGAACGGCCCCACCGGTTATTGTCCGAGTTTGACATCGGCCAGTCTATATTCAACACCTGCCACAAAATTAGAACGGGTTGAGTCTCTCTACGGAGGGACAAGGTCGCTGTATTGTAAATCGCCTCCACTCACTAGAGCCAATTTAAACCGCTCGCAGTCGGTTTACACGAAAAACAGCGGCGGCCCTCGGGACATCCTCCCGAAGCCCGGTCCCCTGGTCCCCACGCAGTCGCTACATTCGATAAAACTGAACCAACAAACGCACGTTCATAACCAGAACTTGCAAAATCAGCTCCAACAGCGCTCCGAAAGCGTGTATGGGGTGAGGGGAAACCCGCCGGGGAGCCTCTCGCGGCCCGAAACCGAGAAAAGCGCCTATCCCAGTAAAAATAGCGCGCCGAATAGTGAAGAGAGCAAAAAGAACTCGAAGATAAACAGGCCGGAGTCGATGTACGGGATTATGAGTTTGCAGCGTAGGGGACAGTCGGCCCAGTCGGACGACTCGAGCTACGGGTCCTATCATGGACCAGCTCCGAGGAACAACACGTATCATTCCAACACTTCGGGTTTCCAGAATAAGGCCGGAAGCGTGGCGAGTTATGTGAGCAGGAACGCCGGGTCCGCGGAGGGGAGACCGCCCCCTTCGGCAAACCAGCTCCTCACCACCACAGGAAACCCGACCAATTCGTACCATCACCACCAACACTCGCCCTCGTCTCAATATTAA
Tribolium Transcript: 3' UTR:
TACTGCGAGCATTGAGATTACCGTTTGTACAGCATTTACAAGTTTCGATGTATCCAAAGAATAGCCGATAGTTTTTTAGATTAGGCTTACGTGTAAGTAGGTATATGTATTATGGTTATACTTTACGATAGGTACATAGAAAAGTGTTCTTGTTTAGCCAGTAGTTTATTTATAAACAGGACTGAAACCGACATGTCTCCAATAAAACGTTTTTGAAAA
Tribolium Protein:
MAEERVNLSSEPSIDYHIVTLFEKLEALRNDKPNSQFTSKLSIKVEVRTLEFWRSVISECIASFVYVFVVCGAKAGAFDGGSATTSLGVLSTALAAGFAMISLTQCFGHISGAHINPAVSLAMGITKNISLLRTTMFIVAQCGGAIAGAAFLYGVSIPRTQEDLPMSLPIKDGPWERFGNEFILTFVVVLAYFVSMDAQRRWMGMSAITIGATYSACSFVSPMPHLNPARSLGPSFVLKKWDNHWVYWLGPIIGGVAAGLIHEYIFNSKRSKRQNSSSDGDSSSIQSDEDTYDDLDKPNAPKYQESTYNSYRAGAPNGPTGYCPSLTSASLYSTPATKLERVESLYGGTRSLYCKSPPLTANLNRSQSVYTKNSGGPRDILPKPGPLVPTQSLHSIKLNQQTHVHNQNLQNQLQQRSESVYGVRGNPPGSLSRPETEKSAYPSKNSAPNSEESKKNSKINRPESMYGIMSLQRRGQSAQSDDSSYGSYHGPAPRNNTYHSNTSGFQNKAGSVASYVSRNAGSAEGRPPPSANQLLTTTGNPTNSYHHHQHSPSSQY
Forward Primer 1:
TCGCAGTCGGTTTACACGAA
Reverse Primer 1:
GGCGCGCTATTTTTACTGGG
Forward Primer 2:
CTCCGAGTGCATAGCGTCTT
Reverse Primer 2:
CATGCCCATCCACCTCCTTT
Gel Picture:
Cloned Fragment:
CTCCGAGTGCATAGCGTCTTTTGTTTACGTGTTTGTGGTGTGCGGTGCAAAGGCAGGTGCTTTTGACGGAGGTTCGGCAACTACCTCTTTGGGAGTTCTTTCAACAGCACTGGCAGCTGGCTTTGCAATGATATCGTTAACACAGTGCTTCGGACACATCTCTGGGGCGCACATAAACCCGGCCGTGAGTCTGGCCATGGGCATTACGAAGAATATTTCGCTTCTGCGGACGACAATGTTCATTGTCGCACAATGCGGGGGTGCGATCGCCGGCGCCGCTTTCCTCTACGGCGTATCCATTCCGCGAACTCAGGAGGATCTCCCGATGTCCCTGCCGATAAAAGATGGGCCTTGGGAGAGGTTCGGGAACGAGTTTATTCTGACTTTCGTGGTGGTTTTGGCCTATTTCGTCTCAATGGACGCCCAAAGGAGGTGGATGGGCATG
Injection concentration:
1ug/ul
Number Injected:
20
Number Survived to Adult:
14
Number w/ RNAi Phenotype:
0
Percent Penetrance:
0%
Assay Description:
N/A
RNAi Phenotype Description:
There was no observable phenotype
RNAi Phenotype/Assay Images:
Assay Data:
N/A
Discussion:
The results showed a 30% mortality rate in larva injected with the RNAi. Those that survived into adulthood had no mortality rate due to RNAi and had no observable phenotype. The phenotype of Tribolium Castaneum differs from the loss of function phenotype in Drosophila, who showed bigger brains and loss of sensory hairs.
These results neither confirm nor deny conserved function in the beetles. The lethality in the larva suggest the gene is involved in something important but does not indicate what. The gene could be involved in CNS development or it could be involved in something else. The survival of the adults suggests a few things. The first one is there is a specific stage in early development the gene affects; if the beetles were not injected early enough there was no effect on the beetles. The next logical point is that the gene is important in early development but is not involved in beetles during their development into adulthood. Further understanding of the function of BIB will illuminate the complexities of insects during their process of changing into adults and how that differs from initial developmental processes.
Notebook:
Annotated Genbank File:
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