Jakob_PS2 - Problem
Set
2

 
 


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Unformatted text preview: Problem
Set
2

 
 
 MCB
104
–
Section
105/106
 1. What
is
the
key
distinction
between
a
cell
strain
and
a
cell
line?
What
are
the
 advantages
and
disadvantages
of
using
each
for
experiments?
 
 
 
 2. How
can
you
experimentally
change
the
sub‐cellular
localization
of
a
protein?
 
 
 
 3. Consider
the
accompanying
pedigree
of
a
rare
autosomal
recessive
disease,
PKU.
 (Griffiths
et
al.,
Chapter
2,
Q:
42).
 
 
 
 
 
 
 
 
 a. List
the
genotypes
of
as
many
of
the
family
members
as
possible.
 
 b. If
persons
A
and
B
marry,
what
is
the
probability
that
their
first
child
will
have
PKU?
 c. If
their
first
child
is
normal,
what
is
the
probability
that
their
second
child
will
have
 PKU?
 d. If
their
first
child
has
the
disease,
what
is
the
probability
that
their
second
child
will
 be
unaffected?
 (Assume
that
all
people
marrying
into
the
pedigree
lack
the
abnormal
allele.)
 4. One
of
the
flow
cytometers
on
the
4th
floor
of
LSA
has
the
excitation
lasers
and
 emission
filters
given
in
the
table
below.
Each
emission
filter
can
only
be
used
with
 the
excitation
laser
to
its
left.
The
excitation
and
emission
spectra
of
five
fluorescent
 proteins
are
also
provided.

 
 Flow
Cytometer
 Excitation
Laser
 488
+/‐
20
 Emission
Filters
 780
+/‐
30
 695
+/‐
20
 575
+/‐
13
 530
+/‐
15
 525
+/‐
25
 450
+/‐
25
 780
+/‐
30
 660
+/‐
10
 405
+/‐
20
 633
+/‐
20
 
 Fluorescent
Proteins
 Left
to
right:
CFP,
GFP,
YFP,
Tomato,
Cherry
 
 
 Using
the
information
above,
answer
the
following
questions.
Assume
that
at
least
50%
 absorption
and
emission
is
required
to
detect
a
protein.

 
 a. Which
of
these
five
fluorescent
proteins
can
the
flow
cytometer
detect?
 
 
 b. If
you
wanted
to
observe
two
proteins
in
a
cell
at
the
same
time,
what
would
be
two
 good
fluorescent
markers
to
use
on
this
flow
cytometer
(Hint:
each
filter
should
 detect
only
one
fluorescent
protein)?
 
 
 c. If
two
different
fluorescent
proteins
are
very
close
together
in
space,
the
emission
 energy
of
one
protein
can
be
accepted
as
excitation
energy
by
the
neighboring
 protein
through
a
process
called
“fluorescence
resonance
energy
transfer”
or
FRET.
 If
this
happens,
the
energy
emitted
from
the
first
fluorophore
is
never
detected.
 Name
two
of
the
above
proteins
that
would
make
a
good
FRET
pair.
Which
 excitation
laser
and
which
emission
filter
would
you
use
to
detect
the
interaction?
 
 
 CFP:
410‐470/460‐520
 GFP:
450‐500/500‐530
 YFP:
500‐525/520‐550
 Tomato:
510‐570/560‐630
 Cherry:
540‐600/590‐650
 ...
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This note was uploaded on 04/05/2010 for the course MCB 104 taught by Professor Urnov during the Spring '09 term at University of California, Berkeley.

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