pGBKT7 - Genes & Developmentgenesdev.cshlp.org/content/suppl/2015/06/10/29.11.1188.DC1/... ·...

L- T - L- T -A-pGAD/pGBKT7pGAD-Fob1/pGBKT7

pGAD-N-Fob1/pGBKT7pGAD-N-Fob1/pGBK-RPA2

pGAD-Fob1/pGBKT7-RPA2

pGAD/pGBKT7-RPA2

0

pGAD

/pG

BT

pGAD

N-F

ob1/

pGB

KT7

RPA

2

pGAD

-N-F

ob1/

pGB

KT7

pGAD

Fob1

/pG

BK

T7R

PA23.0

2.0

1.0

ββga

lact

osid

ase

units

B

A

wt- C- Fob1m3-C-fob1

1 2 3 4

Inte

nsity

(arb

itrar

yun

its)

8000

6000

4000

2000

0m3-C-Fob132P

N- Fob1-GST GSTIn 1 2 3 4 In 1 2 3 4

In 1 2 3 4N- Fob1-GST GST

wt-C-Fob132P

In 1 2 3 4

i ii

C

D

Choudhury et al. Fig.S2

pGAD-NFob1/ pGBKT7

pGAD-NFob1/ pGBKT7 RPA2

pGAD-Fob1E373V/ pGBKT7

pGAD-Fob1E373V/ pGBKT7 RPA2

pGAD-Fob1T504A-S519A/ pGBKT7 RPA2

pGAD-Fob1T504D-S519D/ pGBKT7 RPA2

(X 10-1pmole)

L- T -A-L- T -

pGAD-Fob1-K454T/pGBT9 (I)pGAD-Fob1-K454T/pGBT9-Fob1(II)

pGAD-Fob1-K454T/pGBT9-NFob1(III)pGAD-Fob1/pGBT9-Fob1(IV)

pGAD-NFob1/pGBT9-NFob1(V)

0

4

8

12

16

ββga

lact

osid

ase

activ

ity

I II III IV V

A

B


L- T - L- T -A-

pGBT9/pGADFob1 (I)

pGBT9-Fob1/pGADFob1 (II)pGBT9/pGADFob1(373) (III)pGBT9-Fob1(373)/pGADFob1(373) (IV)pGBT9/pGAD-N-Fob1(V)

pGBT9-NFob1/pGAD-NFob1(VI)pGBT9/pGAD-NFob1(373)(VII)pGBT9-NFob1(373)/pGAD-NFob1(373)(VIII)

18

14

10

6

20

ββ-ga

lact

ivity

I II III IV V VI VII VIII

integrat

ed

Free

plasmid

WT sir2∆∆Fob1

sir2∆∆Fob1E

373V

sir2∆∆fob1∆∆

sir2∆∆NFob1E

373V

C

0

20

40

60

80

100

YPK9E373VE373V∆∆C1E373V∆∆C2E373V∆∆C3E373V∆∆C4

0 20 40 60generations

%su

rviv

al

A

B

D


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