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Cristina Al-Jageta

Friedrich Lab

Evolution of the Methuselah Gene Family in Schizophoran Flies

Throughout this semester, the goal of our research team has been to correctly identify the
relationship genes in the Methuselah gene family in different Schizophoran fly species. The
Methuselah gene is one of the aging genes known in Drosophila melanogaster. The Methuselah
gene itself is just a few million years old. Specifically, our research team focused on the
Methuselah gene family members 1, 5, 14, and 15, which were the oldest Methuselah genes we
could identify.

My research team and I went about searching for the Methuselah gene family content in
the newly available fly genomes. More specifically, we used the Drosophila Melanogaster
Methuselah 1, 5, 14, and 15 sequences as a starting point to find orthologs of Methuselah in other
species. The other species we focused on included Stomoxys calcitrans, Glossina morsitans,
Ceratitis capitata, Musca domestica, Tribolium castaneum, and Drosophila virilis.

Each member of the research team was assigned a specific species within the
Schizophoran fly family. I was responsible for the species Glossina morsitans. Glossina
morsitans is a biting fly that causes sleeping sickness in humans. It is related to the housefly and
very common in Africa. To begin, I utilized several databases including Flybase and NCBI to
find ortholog sequences of my species. To ensure accurate results, I reverse blasted the
sequences of the discovered orthologs using NCBI. Reverse blasting the ortholog sequences was
critical for reliable prediction of gene function in these newly sequenced genomes. Following the
reverse blasts, I used Muscle and Clustal Alignments to find incomplete sequences and replace
them with more complete ones. Then, using the muscle alignment and G-blocks databases I
received the alignment formation necessary for a program, MEGA, which helps constructs
phylogenetic trees. Using the MEGA software, I was able to construct a phylogenetic tree using
all the ortholog sequences through MEGA.

Attached is the phylogenetic tree I constructed for my specific species, Glossina

morsitans. Through my analysis of the genomes for this species, I have found that there are 8
Methuselah orthologs present in the Glossina morsitans species. Using this phylogenetic tree and
the genome of my species, the members of my research team and I were able to construct a
phylogenetic tree containing all the species in question. I have also attached the phylogenetic tree
we constructed. Methuselah genes 1, 5, 14, and 15 are conserved as 1:1 orthologs. This result
was what we hypothesized due to the fact that they are the oldest and most conserved
Methuselah genes.

At this point in time, we are interested in find the degree of conservation within the
Methuselah orthologs. The members of my research group and I are constructing separate
alignments of Mthl 1, 5, 14, and 15 orthologs our species. I will be constructing this alignment
for Glossina morsitans using the alignment database Muscle.