This document outlines three research positions held by the author between 2013-2015. The first was in an influenza virus laboratory focusing on protein-protein interactions between host SUMO proteins and viral proteins. Tasks involved assays to study interactions between SUMO-1 and influenza NS1 protein. The second position was in a Mycobacterium tuberculosis laboratory, where the author generated gene knockouts in Mtb and BCG strains to study the effect on growth. The third was a brief summer internship assisting with cloning a vector to infect neuronal cells and study glycinergic neurotransmission.

1. Job
Descriptions
Mentor:
Dr.
German
Rosas-­‐Acosta
Duration:
October
2014
-­‐
August
2015
This
was
an
influenza
virus
laboratory
that
focused
on
the
elucidations
of
viral
and
host
protein
interactions,
primarily
how
host
factor
SUMO
(Small
ubiquitin-­‐like
modifier)
would
SUMO-­‐ylate
viral
factors
and
either
optimize
infection
or
slow
it.
My
task
while
I
was
in
this
lab
involved
a
lot
of
protein-­‐protein
interaction
assays
between
SUMO-­‐1
(a
homologue
of
SUMO)
and
how
it
interacts
with
influenza
viral
protein
NS1
(Which
is
a
protein
that
blocks
host
interferon
response).
In
the
past
it
had
been
elucidated
that
this
interaction
occurs
in
an
enzyme
driven
SUMO-­‐cycle
cascade,
however
recent
hypothesis
state
there
is
an
alternate
interaction
that
occurs
between
hydrophobic
motifs
in
each
of
the
respective
proteins.
Many
pulldown
assays
were
done
to
measure
to
observe
this
interaction
and
quantify
it's
strength
via
autoradiography
methods
and
western
blot
analysis.
Mentor:
Dr.
Hugues
Ouellet
Duration:
September
2013
–
October
2014
This
lab
was
a
lab
focused
on
the
pathobiology
of
Mycobacterium
tuberculosis.
I
was
responsible
for
the
creation
of
gene
knockouts
in
Mtb
and
on
the
attenuated
strain
Bacille
Calmette
Guerin
(BCG)
to
observe
how
the
gene
glbN
affected
the
growth
and
stability
of
the
mycobacterium
in
vitro.
glbN
is
a
gene
involved
in
the
detoxification
of
host
macrophage-­‐generated
nitric
oxide.
I
was
responsible
for
making
the
glbN
knock
out
(∆glbN)
from
scratch
including
the
creation
of
a
recombinant
vector
and
creation
of
clones,
the
methods
employed
were:
PCR,
digestion,
ligation,
transformation,
clone
screening,
and
the
packaging
of
the
vector
into
a
mycobacterial
phage
via
transduction
in
both
BCG
and
Mtb
strains.
Only
Dr.
Hugues
Ouellet
had
access
to
BSL3
and
so
my
job
was
to
overlook
and
monitor
the
growth
of
the
new
BCG
knockouts.
The
next
phase
involved
making
growth
curves
to
compare
the
growth
of
both
wild
type
and
∆glbN
strains
in
different
growth
mediums
and
track
it's
discrepancies
via
a
spectrophotometer.
Cells
were
collected
at
different
times
(lagging,
exponential
and
plateau)
to
be
lysed
and
ran
western
blot
analysis
to
observe
the
pattern
of
expression
of
glbN
in
the
wild
type
cells.
This
project
revealed
many
interesting
things
along
the
way
like
the
consideration
of
glycerol
and
nitrogen
sources
from
different
growth
mediums
and
how
they
affected
growth
patterns,
when
I
left
the
lab
we
were
starting
to
look
into
these
questions.
Along
the
way
I
trained
a
new
member
to
the
lab
who
was
hired
to
assist
me
in
this
project.
Mentor:
Dr.
Manuel
Miranda-­‐Arango
Duration:
May
2013
–
August
2013
(Summer
internship)
This
was
the
first
summer
I
was
exposed
to
research
and
it
was
for
a
short
period
of
two
months.
I
wasn't
responsible
for
any
particular
project
but
this
experience
was
transformative
in
teaching
me
about
a
research
environment.
I
did
however
assist
a
PhD
student
in
creating
a
vector
that
was
going
to
be
packaged
in
a
lentivirus
to
infect
neuronal
cells.
This
was
a
neurobiology
lab
that
focused
on
glycinergic
neurotransmission
and
investigated
how
failure
in
its
reuptake
could
lead
to
various
neurological
conditions
ranging
from
schizophrenia
to
hyperekplexia.
This
is
where
I
learned
basic
cloning
techniques
like
PCR
and
bacterial
transformations.