Download Document

Yeast two-hybrid system:
a genetic assay for detecting protein-protein interactions
Suppose we have two proteins...
X
Y
and the question,
do these two protein bind each other?
One way of answering this question is to
use the yeast two hybrid method.
To understand the method we must first
consider how gene expression is regulated in yeast......
Yeast two-hybrid system:
a genetic assay for detecting protein-protein interactions
Regulation of gene expression in yeast
t
r
a
n
s
c
r
i
p
t
i
o
n
a
c
t
i
v
a
t
o
r
a
c
t
i
v
a
t
i
o
n
d
o
m
a
i
n
t
r
a
n
s
c
r
i
p
t
i
o
n
m
a
c
h
i
n
e
r
y
D
N
A
b
i
n
d
i
n
g
d
o
m
a
i
n
U
A
S
(
u
p
s
t
r
e
a
m
a
c
t
i
v
a
t
i
o
n
s
e
q
u
e
n
c
e
)
g
e
n
e
and now back to the question...
Yeast two-hybrid system:
a genetic assay for detecting protein-protein interactions
Regulation of gene expression in yeast
t
r
a
n
s
c
r
i
p
t
i
o
n
a
c
t
i
v
a
t
o
r
a
c
t
i
v
a
t
i
o
n
d
o
m
a
i
n
D
N
A
b
i
n
d
i
n
g
d
o
m
a
i
n
U
A
S
(
u
p
s
t
r
e
a
m
a
c
t
i
v
a
t
i
o
n
s
e
q
u
e
n
c
e
)
t
r
a
n
s
c
r
i
p
t
i
o
n
m
a
c
h
i
n
e
r
y
D
othese
tw
oprotein
bind?
g
e
n
e
Y
X
We start with yeast possessing a reporter gene (a gene
making a product that is easy to detect).
And now we introduce genes coding for two hybrid
proteins.....
r
e
p
o
r
t
e
r
g
e
n
e
Yeast two-hybrid system:
a genetic assay for detecting protein-protein interactions
Regulation of gene expression in yeast
t
r
a
n
s
c
r
i
p
t
i
o
n
a
c
t
i
v
a
t
o
r
a
c
t
i
v
a
t
i
o
n
d
o
m
a
i
n
t
r
a
n
s
c
r
i
p
t
i
o
n
m
a
c
h
i
n
e
r
y
D
N
A
b
i
n
d
i
n
g
d
o
m
a
i
n
g
e
n
e
U
A
S
(
u
p
s
t
r
e
a
m
a
c
t
i
v
a
t
i
o
n
s
e
q
u
e
n
c
e
)
Y
o
u
r
t
w
o
h
y
b
r
i
d
p
r
o
t
e
i
n
s
b
i
n
d
!
yes, we have expression of
the reporter indicating that
the proteins bind!
Y
t
h
u
s
f
o
r
m
i
n
g
a
f
u
n
c
t
i
o
n
a
l
t
r
a
n
s
c
r
i
p
t
i
o
n
a
c
t
i
v
a
t
o
r
.
.
. X
Y
X
X
X
Do these
proteins
bind?
Y
do we get expression of the reporter?
r
e
p
o
r
t
e
r
g
e
n
e
e
n
d
o
f
s
l
i
d
e
s
h
o
w