Rare Diseases Symptoms Automatic Extraction
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Discovery of protein interactions using parallel analysis of translated ORFs (PLATO).
[inclusion body myositis]
Parallel
analysis
of
translated
open
reading
frames
(
ORFs
)
(
PLATO
)
can
be
used
for
the
unbiased
discovery
of
interactions
between
full-length
proteins
encoded
by
a
library
of
'
prey
'
ORFs
and
surface-immobilized
'
bait
'
antibodies
,
polypeptides
or
small
-molecular-weight
compounds
.
PLATO
uses
ribosome
display
(
RD
)
to
link
ORF-derived
mRNA
molecules
to
the
proteins
they
encode
,
and
recovered
mRNA
from
affinity
enrichment
is
subjected
to
analysis
using
massively
parallel
DNA
sequencing
.
Compared
with
alternative
in
vitro
methods
,
PLATO
provides
several
advantages
including
library
size
and
cost
.
A
unique
advantage
of
PLATO
is
that
an
alternative
reverse
transcription-quantitative
PCR
(
RT-qPCR
)
protocol
can
be
used
to
test
binding
of
specific
,
individual
proteins
.
To
illustrate
a
typical
experimental
workflow
,
we
demonstrate
PLATO
for
the
identification
of
the
immune
target
of
serum
antibodies
from
patients
with
inclusion
body
myositis
(
IBM
)
.
Beginning
with
an
ORFeome
library
in
an
RD
vector
,
the
protocol
can
produce
samples
for
deep
sequencing
or
RT-qPCR
within
4
d
.
