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Table 2 Analysis of fluorescent and non-fluorescent mutants carrying single amino acid substitutions

From: The effect of amino acid deletions and substitutions in the longest loop of GFP

WT aa

AS

Hydrophobic aa

Hydrophilic aa

Acid

Basic

Stop

  

W

F

P

M

L

I

V

A

G

N

Q

S

T

C

Y

D

E

R

K

H

X

F131

B

  

1

 

1

1

 

2*

  

1

 

1

  

1

  

1

1

 

K132

E

1

    

1

 

1

1

       

1

2

   

E133

E

  

1

 

2

 

1

3

1

1

 

1

1

  

1

 

3

   

D134

E

 

1

        

1

2

2

    

2

   

G135

B

    

1

1*

1*

 

1

  

1*

 

1

1

      

N136

E

 

1

4

1

1

 

4

4

  

1

 

2

    

1

1

1

 

I137

B

    

4

 

2

 

1

  

1*

   

1

2

2

  

1

L138

B

 

1

 

2

2

1

  

1*

    

1

   

1

   

G139

E

  

1*

 

2

  

1

1

1

2

2

1

1

    

2

1

 

H140

E

  

1

  

1*

   

1

    

1

  

2

2

  

K141

E

  

2*

1

   

1

 

1

  

1

    

1

 

1

 

L142

B

1

    

1

1

 

2

1

 

1

   

1

     
  1. Mutants displaying a green-fluorescent phenotype to the naked eye are shown in bold numbers. Mutants displaying a white phenotype are shown in italic numbers. Mutants displaying a pale green-fluorescent phenotype are shown in underlined numbers. Those mutants labeled with asterisk turned pale green-fluorescent after prolonged growing at room temperature. The number represents the number of times that each mutant was found. In this table, empty spaces represent wild-type amino acids. B means buried amino acid; E, solvent exposed amino acid. During screening of fluorescent mutants, seven clones corresponded to wt sgGFP generated with the mixtures of degenerated oligonucleotides.