      

                                SEQUENCE LISTING

<110> Kleinfeld, Alan Marc
	Huber, Andrew Henry
	Kampf, James Patrick
	Kwan, Thomas
	Zhu, Baolong

<120> Development and Use of Cysteine -Labeled Fluorescent Probes of 	Unbound Analytes

<130> FFASC.072PR


<160> 5

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 399
<212> DNA
<213> Rattus norvegicus

<220> 
<221> CDS            
<222> (1)...(399)
<223> wild-type rat intestinal fatty acid binding protein cDNA sequence


<400> 1
atg gca ttt gat ggc act tgg aaa gta tac cgg aat gag aac tat gaa   48
Met Ala Phe Asp Gly Thr Trp Lys Val Tyr Arg Asn Glu Asn Tyr Glu
 1               5                   10                  15
 
aag ttc atg gag aaa atg ggc att aac gtg gtg aag agg aag ctt gga   96
Lys Phe Met Glu Lys Met Gly Ile Asn Val Val Lys Arg Lys Leu Gly
             20                  25                  30
 
gct cat gac aac ttg aaa ctg acg atc aca cag gaa gga aat aaa ttc   144
Ala His Asp Asn Leu Lys Leu Thr Ile Thr Gln Glu Gly Asn Lys Phe
         35                  40                  45
 
aca gtc aaa gaa tca agc aac ttc cga aac att gat gtt gtg ttt gaa   192
Thr Val Lys Glu Ser Ser Asn Phe Arg Asn Ile Asp Val Val Phe Glu
     50                  55                  60
 
ctc ggc gtc gac ttt gcc tat agt cta gca gat gga aca gaa ctc act   240
Leu Gly Val Asp Phe Ala Tyr Ser Leu Ala Asp Gly Thr Glu Leu Thr
 65                  70                  75                  80
 
ggg acc ttg acc atg gag gga aat aaa ctt gtt gga aaa ttc aaa cgt   288
Gly Thr Leu Thr Met Glu Gly Asn Lys Leu Val Gly Lys Phe Lys Arg
                 85                  90                  95
 
gta gac aat gga aag gag ctg att gct gtc cga gag att tct ggt aac   336
Val Asp Asn Gly Lys Glu Leu Ile Ala Val Arg Glu Ile Ser Gly Asn
            100                 105                 110
 
gaa cta atc caa acc tac aca tat gaa gga gtg gag gcc aag cgc atc   384
Glu Leu Ile Gln Thr Tyr Thr Tyr Glu Gly Val Glu Ala Lys Arg Ile
        115                 120                 125
 
ttt aag aag gaa tag                                               399
Phe Lys Lys Glu  *
    130
 

<210> 2
<211> 131
<212> PRT
<213> Rattus norvegicus

<220> 
<221> PEPTIDE        
<222> (1)...(131)
<223> wild-type rat intestinal fatty acid binding
      protein

<220> 
<221> ACETYLATION    
<222> (1)...(1)

<400> 2
Ala Phe Asp Gly Thr Trp Lys Val Asp Arg Asn Glu Asn Tyr Glu Lys
 1               5                  10                  15      
Phe Met Glu Lys Met Gly Ile Asn Val Val Lys Arg Lys Leu Gly Ala
            20                  25                  30          
His Asp Asn Leu Lys Leu Thr Ile Thr Gln Glu Gly Asn Lys Phe Thr
        35                  40                  45              
Val Lys Glu Ser Ser Asn Phe Arg Asn Ile Asp Val Val Phe Glu Leu
    50                  55                  60                  
Gly Val Asp Phe Ala Tyr Ser Leu Ala Asp Gly Thr Glu Leu Thr Gly
65                  70                  75                  80  
Thr Trp Thr Met Glu Gly Asn Lys Leu Val Gly Lys Phe Lys Arg Val
                85                  90                  95      
Asp Asn Gly Lys Glu Leu Ile Ala Val Arg Glu Ile Ser Gly Asn Glu
            100                 105                 110         
Leu Ile Gln Thr Tyr Thr Tyr Glu Gly Val Glu Ala Lys Arg Ile Phe
        115                 120                 125             
Lys Lys Glu
    130     


<210> 3
<211> 426
<212> DNA
<213> Rattus norvegicus

<220> 
<221> mutation       
<222> (1)...(426)
<223> rat intestinal fatty acid binding protein DNA
      sequence coding for substitution of alanine for
      leucine at position 72

<223> 3' terminus codes for a 6his tag

<400> 3
atggcatttg atggcacttg gaaagtagac cggaatgaga actatgaaaa gttcatggag 60
aaaatgggca ttaacgtggt gaagaggaag cttggagctc atgacaactt gaaactgacg 120
atcacacagg aaggaaataa attcacagtc aaagaatcaa gcaacttccg aaacattgat 180
gttgtgtttg aactcggcgt cgactttgcc tatagtgctg cagatggaac agaactcacc 240
ggtacctgga caatggaggg aaataaactt gttggaaagt ttaaacgtgt agacaatgga 300
aaggagctga ttgctgtccg agagatttct ggtaacgaac taatccagac ctacacatat 360
gaaggagtgg aggccaagcg gatctttaag aaggaccgcg gtcatcatca ccatcatcac 420
tagtaa                                                            426

<210> 4
<211> 139
<212> PRT
<213> Rattus norvegicus

<220> 
<221> MUTAGEN        
<222> (72)...(72)
<223> rat intestinal fatty acid binding protein with
      alanine substitution for leucine at position 72 and Glu 131 to Asp 	subsitition.

<223> COOH-terminal affinity tag comprising Arg132, Gly133 and 6 histidines

<400> 4
Ala Phe Asp Gly Thr Trp Lys Val Asp Arg Asn Glu Asn Tyr Glu Lys
 1               5                  10                  15      
Phe Met Glu Lys Met Gly Ile Asn Val Val Lys Arg Lys Leu Gly Ala
            20                  25                  30          
His Asp Asn Leu Lys Leu Thr Ile Thr Gln Glu Gly Asn Lys Phe Thr
        35                  40                  45              
Val Lys Glu Ser Ser Asn Phe Arg Asn Ile Asp Val Val Phe Glu Leu
    50                  55                  60                  
Gly Val Asp Phe Ala Tyr Ser Ala Ala Asp Gly Thr Glu Leu Thr Gly
65                  70                  75                  80  
Thr Trp Thr Met Glu Gly Asn Lys Leu Val Gly Lys Phe Lys Arg Val
                85                  90                  95      
Asp Asn Gly Lys Glu Leu Ile Ala Val Arg Glu Ile Ser Gly Asn Glu
            100                 105                 110         
Leu Ile Gln Thr Tyr Thr Tyr Glu Gly Val Glu Ala Lys Arg Ile Phe
        115                 120                 125             
Lys Lys Asp Arg Gly His His His His His His
    130                 135                 


<210> 5
<211> 139
<212> PRT
<213> Rattus norvegicus

<220> 
<221> MUTAGEN        
<222> (131)...(131)
<223> rat intestinal fatty acid binding protein with Glu 131 to Asp 	subsitition.

<223> COOH-terminal affinity tag comprising Arg132, Gly133 and 6 histidines

<400> 5

Ala Phe Asp Gly Thr Trp Lys Val Asp Arg Asn Glu Asn Tyr Glu Lys
 1               5                  10                  15      
Phe Met Glu Lys Met Gly Ile Asn Val Val Lys Arg Lys Leu Gly Ala
            20                  25                  30          
His Asp Asn Leu Lys Leu Thr Ile Thr Gln Glu Gly Asn Lys Phe Thr
        35                  40                  45              
Val Lys Glu Ser Ser Asn Phe Arg Asn Ile Asp Val Val Phe Glu Leu
    50                  55                  60                  
Gly Val Asp Phe Ala Tyr Ser Leu Ala Asp Gly Thr Glu Leu Thr Gly
65                  70                  75                  80  
Thr Trp Thr Met Glu Gly Asn Lys Leu Val Gly Lys Phe Lys Arg Val
                85                  90                  95      
Asp Asn Gly Lys Glu Leu Ile Ala Val Arg Glu Ile Ser Gly Asn Glu
            100                 105                 110         
Leu Ile Gln Thr Tyr Thr Tyr Glu Gly Val Glu Ala Lys Arg Ile Phe
        115                 120                 125             
Lys Lys Asp Arg Gly His His His His His His
    130                 135                 









					
