                     SEQUENCE LISTING

<110>  BioMarin Pharmaceutical Inc.
       Foehr, Erik Damien
       Zhao, Bin
       Santamaria, Carlos Fabricio

<120>  Assays for detection of phenylalanine ammonia-lyase and 
       antibodies to phenylalanine ammonia-lyase

<130>  11808-029-228

<150>  US 61/084,961
<151>  2008-07-30

<160>  31    

<170>  PatentIn version 3.4

<210>  1
<211>  1710
<212>  DNA
<213>  Nostoc punctiforme

<400>  1
atgaatataa catctctaca acagaacata acgcgttctt ggcaaatacc tttcactaat     60

agttcagatt caatcgtaac tgtaggcgat cgcaatctga caatcgacga ggttgtaaat    120

gttgctcgtc atggaacaca ggtgcgctta actgataatg cagatgtcat tcggggtgtt    180

caagcatctt gtgattacat taacaatgca gtcgaaacag cacagccaat ttacggggtg    240

acatctggct ttggcggtat ggcagatgtt gtcatctctc gcgaacaagc agcggaactt    300

cagactaatt taatttggtt tctgaaatcc ggcgcaggaa acaaattatc gttagcagac    360

gtgcgtgcag ctatgctctt acgtgcaaat tcacatttgt atggtgcgtc tggtatacga    420

ctcgaactta ttcagcggat tgaaactttc ctcaacgctg gcgtgacacc ccatgtctat    480

gagtttggct ctatcggtgc tagcggcgat ttggtgccat tatcctacat tactggggca    540

ctaatcggtc tagatcctag ctttacagtt gacttcgacg gtaaagaaat ggatgccgtt    600

acagccttgt ctcgtttggg tttgccaaag ttgcaattgc aaccgaaaga aggtttagca    660

atgatgaatg gcacctcagt catgacaggt attgcagcta actgtgtgta cgatgcgaaa    720

gttttgctcg ctctgacaat gggtgtacac gccttagcca tccaaggttt atacggaacg    780

aatcaatctt tccacccgtt tattcatcag tgcaagccac atcccggtca actatggaca    840

gcagatcaaa tgttttctct gctgaaagat tcatctttag ttcgtgaaga gttggatggt    900

aaacacgaat accgtggtaa agatctgata caggatcgtt attctctccg ctgtctggca    960

cagttcatag ggccaatcgt tgatggggta tcagagatta ccaagcaaat cgaggtagaa   1020

atgaactcag tcaccgataa cccattgatt gatgtcgaga accaagttag ttatcacggc   1080

ggcaattttc tcggacagta tgtgggtgtg acaatggatc gcctacgtta ttacataggg   1140

ctattggcca aacacatcga tgtgcagatt gcacttcttg tctcgccaga gtttagcaac   1200

ggcttaccac cctctttagt tggtaatagc gatcgcaaag ttaatatggg actcaaaggt   1260

ttgcaaatca gtggaaactc gattatgcca ctgttgagct tctatggaaa ttccctagcc   1320

gatcgctttc ctacccacgc cgagcaattt aatcaaaata ttaacagcca aggctatatt   1380

tccgcaaatt tgacacgtcg ttccgtagac atatttcaga attatatggc gatcgcgttg   1440

atgtttggag ttcaagctgt tgacctccgc acatataaga tgaaaggtca ttatgatgca   1500

cgtacatgcc tctcacccaa tactgtgcag ttatacacag cagtctgcga ggtagttgga   1560

aagccactaa cgtctgtgcg tccatacatt tggaacgaca acgagcaatg tttagatgag   1620

catattgccc ggatttcagc tgatatcgct ggtggtggtt taattgtgca agcagttgag   1680

catatttttt cgagcttaaa gtcaacgtaa                                    1710

<210>  2
<211>  569
<212>  PRT
<213>  Nostoc punctiforme

<400>  2

Met Asn Ile Thr Ser Leu Gln Gln Asn Ile Thr Arg Ser Trp Gln Ile 
1               	5                   10                  15      

Pro Phe Thr Asn Ser Ser Asp Ser Ile Val Thr Val Gly Asp Arg Asn 
            20                  25                  30          

Leu Thr Ile Asp Glu Val Val Asn Val Ala Arg His Gly Thr Gln Val 
        35                  40                  45              

Arg Leu Thr Asp Asn Ala Asp Val Ile Arg Gly Val Gln Ala Ser Cys 
    50                  55                  60                  

Asp Tyr Ile Asn Asn Ala Val Glu Thr Ala Gln Pro Ile Tyr Gly Val 
65                  70                  75                  80  

Thr Ser Gly Phe Gly Gly Met Ala Asp Val Val Ile Ser Arg Glu Gln 
                85                  90                  95      

Ala Ala Glu Leu Gln Thr Asn Leu Ile Trp Phe Leu Lys Ser Gly Ala 
            100                 105                 110         


Gly Asn Lys Leu Ser Leu Ala Asp Val Arg Ala Ala Met Leu Leu Arg 
        115                 120                 125             

Ala Asn Ser His Leu Tyr Gly Ala Ser Gly Ile Arg Leu Glu Leu Ile 
    130                 135                 140                 

Gln Arg Ile Glu Thr Phe Leu Asn Ala Gly Val Thr Pro His Val Tyr 
145                 150                 155                 160 

Glu Phe Gly Ser Ile Gly Ala Ser Gly Asp Leu Val Pro Leu Ser Tyr 
                165                 170                 175     

Ile Thr Gly Ala Leu Ile Gly Leu Asp Pro Ser Phe Thr Val Asp Phe 
            180                 185                 190         

Asp Gly Lys Glu Met Asp Ala Val Thr Ala Leu Ser Arg Leu Gly Leu 
        195                 200                 205             

Pro Lys Leu Gln Leu Gln Pro Lys Glu Gly Leu Ala Met Met Asn Gly 
    210                 215                 220                 

Thr Ser Val Met Thr Gly Ile Ala Ala Asn Cys Val Tyr Asp Ala Lys 
225                 230                 235                 240 

Val Leu Leu Ala Leu Thr Met Gly Val His Ala Leu Ala Ile Gln Gly 
                245                 250                 255     

Leu Tyr Gly Thr Asn Gln Ser Phe His Pro Phe Ile His Gln Cys Lys 
            260                 265                 270         

Pro His Pro Gly Gln Leu Trp Thr Ala Asp Gln Met Phe Ser Leu Leu 
        275                 280                 285             

Lys Asp Ser Ser Leu Val Arg Glu Glu Leu Asp Gly Lys His Glu Tyr 
    290                 295                 300                 

Arg Gly Lys Asp Leu Ile Gln Asp Arg Tyr Ser Leu Arg Cys Leu Ala 
305                 310                 315                 320 

Gln Phe Ile Gly Pro Ile Val Asp Gly Val Ser Glu Ile Thr Lys Gln 
                325                 330                 335     

Ile Glu Val Glu Met Asn Ser Val Thr Asp Asn Pro Leu Ile Asp Val 
            340                 345                 350         

Glu Asn Gln Val Ser Tyr His Gly Gly Asn Phe Leu Gly Gln Tyr Val 
        355                 360                 365             

Gly Val Thr Met Asp Arg Leu Arg Tyr Tyr Ile Gly Leu Leu Ala Lys 
    370                 375                 380                 

His Ile Asp Val Gln Ile Ala Leu Leu Val Ser Pro Glu Phe Ser Asn 
385                 390                 395                 400 

Gly Leu Pro Pro Ser Leu Val Gly Asn Ser Asp Arg Lys Val Asn Met 
                405                 410                 415     

Gly Leu Lys Gly Leu Gln Ile Ser Gly Asn Ser Ile Met Pro Leu Leu 
            420                 425                 430         

Ser Phe Tyr Gly Asn Ser Leu Ala Asp Arg Phe Pro Thr His Ala Glu 
        435                 440                 445             

Gln Phe Asn Gln Asn Ile Asn Ser Gln Gly Tyr Ile Ser Ala Asn Leu 
    450                 455                 460                 

Thr Arg Arg Ser Val Asp Ile Phe Gln Asn Tyr Met Ala Ile Ala Leu 
465                 470                 475                 480 

Met Phe Gly Val Gln Ala Val Asp Leu Arg Thr Tyr Lys Met Lys Gly 
                485                 490                 495     

His Tyr Asp Ala Arg Thr Cys Leu Ser Pro Asn Thr Val Gln Leu Tyr 
            500                 505                 510         

Thr Ala Val Cys Glu Val Val Gly Lys Pro Leu Thr Ser Val Arg Pro 
        515                 520                 525             

Tyr Ile Trp Asn Asp Asn Glu Gln Cys Leu Asp Glu His Ile Ala Arg 
    530                 535                 540                 

Ile Ser Ala Asp Ile Ala Gly Gly Gly Leu Ile Val Gln Ala Val Glu 
545                 550                 555                 560 

His Ile Phe Ser Ser Leu Lys Ser Thr 
                565                 

<210>  3
<211>  1704
<212>  DNA
<213>  Anabaena variabilis

<400>  3
atgaagacac tatctcaagc acaaagcaaa acctcatctc aacaattttc ttttactgga     60

aattcttctg ccaatgtaat tattggtaat cagaaactca caatcaatga tgttgcaagg    120

gtagcgcgta atggcacctt agtgtcttta accaataaca ctgatatttt gcagggtatt    180

caggcatctt gtgattacat taataatgct gttgaatctg gggaaccaat ttatggagtg    240

acatctggtt ttggcggtat ggccaatgtt gccatatccc gtgaacaagc atctgaactc    300

caaaccaact tagtttggtt cctgaaaaca ggtgcaggga acaaattacc cttggcggat    360

gtgcgcgcag ctatgctctt gcgtgcaaac tctcatatgc gcggtgcatc tggcatcaga    420

ttagaactta tcaagcgtat ggagattttc cttaacgctg gtgtcacacc atatgtgtat    480

gagtttggtt caattggtgc aagtggtgat ttagtgccac tatcctacat tactggttca    540

ctgataggct tagatcccag ttttaaggtt gacttcaacg gtaaagaaat ggatgcgcca    600

acagctctac gtcaactgaa tttgtcaccc ttgacattgt tgccgaagga aggcttggcg    660

atgatgaacg gcacttcagt catgacaggt attgcagcaa actgcgtcta cgatactcaa    720

attttaactg cgatcgctat gggcgttcac gctctagata tccaagcttt aaacggaacc    780

aatcaatcat tccatccatt tatccataat tccaaaccac atcctggtca attatgggca    840

gcagatcaga tgatttcttt gttagccaat tcccagttag ttcgtgatga gttagatggt    900

aaacacgatt atcgtgatca cgagttgatt caagatcgtt actcactccg atgccttccc    960

cagtatttgg ggccaatcgt tgatggaatt tcccagattg ccaaacaaat tgaaatcgaa   1020

atcaactcag tcaccgataa cccactaatt gatgttgata accaagctag ctatcatgga   1080

ggaaatttcc tcggacagta cgtgggtatg ggaatggatc acctgcgtta ctatattggg   1140

ttattggcta aacacctaga tgtgcagatt gccctcctcg cctcaccaga gtttagcaat   1200

ggactaccac catctttatt aggcaaccga gaacgtaaag tcaatatggg actcaaaggt   1260

ctgcaaatat gcggtaactc aattatgcca ctgttgacct tctatggaaa ttccatcgcc   1320

gatcgctttc ctacccatgc agaacaattt aatcagaaca tcaacagtca aggatacact   1380

tcagcgactc tagcccgccg ttctgtggat atcttccaga attatgtggc gatcgctctg   1440

atgtttggag tccaagctgt tgacctccgc acatataaaa agactggtca ttacgatgca   1500

cgcgcctgtc tatcacctgc aactgagcgc ttatattcag cagtccgcca cgtagttgga   1560

caaaaaccaa cttcagatcg cccatatatt tggaatgata atgagcaagg actggatgag   1620

catattgccc ggatttctgc tgatatcgct gctggtggtg tgattgtgca agcagttcaa   1680

gatatcttac cctgcttgca ttaa                                          1704

<210>  4
<211>  567
<212>  PRT
<213>  Anabaena variabilis

<400>  4

Met Lys Thr Leu Ser Gln Ala Gln Ser Lys Thr Ser Ser Gln Gln Phe 
1               5                   10                  15      

Ser Phe Thr Gly Asn Ser Ser Ala Asn Val Ile Ile Gly Asn Gln Lys 
            20                  25                  30          

Leu Thr Ile Asn Asp Val Ala Arg Val Ala Arg Asn Gly Thr Leu Val 
        35                  40                  45              

Ser Leu Thr Asn Asn Thr Asp Ile Leu Gln Gly Ile Gln Ala Ser Cys 
    50                  55                  60                  

Asp Tyr Ile Asn Asn Ala Val Glu Ser Gly Glu Pro Ile Tyr Gly Val 
65                  70                  75                  80  

Thr Ser Gly Phe Gly Gly Met Ala Asn Val Ala Ile Ser Arg Glu Gln 
                85                  90                  95      

Ala Ser Glu Leu Gln Thr Asn Leu Val Trp Phe Leu Lys Thr Gly Ala 
            100                 105                 110         

Gly Asn Lys Leu Pro Leu Ala Asp Val Arg Ala Ala Met Leu Leu Arg 
        115                 120                 125             

Ala Asn Ser His Met Arg Gly Ala Ser Gly Ile Arg Leu Glu Leu Ile 
    130                 135                 140                 

Lys Arg Met Glu Ile Phe Leu Asn Ala Gly Val Thr Pro Tyr Val Tyr 
145                 150                 155                 160 

Glu Phe Gly Ser Ile Gly Ala Ser Gly Asp Leu Val Pro Leu Ser Tyr 
                165                 170                 175     

Ile Thr Gly Ser Leu Ile Gly Leu Asp Pro Ser Phe Lys Val Asp Phe 
            180                 185                 190         

Asn Gly Lys Glu Met Asp Ala Pro Thr Ala Leu Arg Gln Leu Asn Leu 
        195                 200                 205             

Ser Pro Leu Thr Leu Leu Pro Lys Glu Gly Leu Ala Met Met Asn Gly 
    210                 215                 220                 

Thr Ser Val Met Thr Gly Ile Ala Ala Asn Cys Val Tyr Asp Thr Gln 
225                 230                 235                 240 

Ile Leu Thr Ala Ile Ala Met Gly Val His Ala Leu Asp Ile Gln Ala 
                245                 250                 255     

Leu Asn Gly Thr Asn Gln Ser Phe His Pro Phe Ile His Asn Ser Lys 
            260                 265                 270         

Pro His Pro Gly Gln Leu Trp Ala Ala Asp Gln Met Ile Ser Leu Leu 
        275                 280                 285             

Ala Asn Ser Gln Leu Val Arg Asp Glu Leu Asp Gly Lys His Asp Tyr 
    290                 295                 300                 

Arg Asp His Glu Leu Ile Gln Asp Arg Tyr Ser Leu Arg Cys Leu Pro 
305                 310                 315                 320 

Gln Tyr Leu Gly Pro Ile Val Asp Gly Ile Ser Gln Ile Ala Lys Gln 
                325                 330                 335     

Ile Glu Ile Glu Ile Asn Ser Val Thr Asp Asn Pro Leu Ile Asp Val 
            340                 345                 350         

Asp Asn Gln Ala Ser Tyr His Gly Gly Asn Phe Leu Gly Gln Tyr Val 
        355                 360                 365             

Gly Met Gly Met Asp His Leu Arg Tyr Tyr Ile Gly Leu Leu Ala Lys 
    370                 375                 380                 

His Leu Asp Val Gln Ile Ala Leu Leu Ala Ser Pro Glu Phe Ser Asn 
385                 390                 395                 400 

Gly Leu Pro Pro Ser Leu Leu Gly Asn Arg Glu Arg Lys Val Asn Met 
                405                 410                 415     

Gly Leu Lys Gly Leu Gln Ile Cys Gly Asn Ser Ile Met Pro Leu Leu 
            420                 425                 430         

Thr Phe Tyr Gly Asn Ser Ile Ala Asp Arg Phe Pro Thr His Ala Glu 
        435                 440                 445             

Gln Phe Asn Gln Asn Ile Asn Ser Gln Gly Tyr Thr Ser Ala Thr Leu 
    450                 455                 460                 

Ala Arg Arg Ser Val Asp Ile Phe Gln Asn Tyr Val Ala Ile Ala Leu 
465                 470                 475                 480 

Met Phe Gly Val Gln Ala Val Asp Leu Arg Thr Tyr Lys Lys Thr Gly 
                485                 490                 495     

His Tyr Asp Ala Arg Ala Cys Leu Ser Pro Ala Thr Glu Arg Leu Tyr 
            500                 505                 510         

Ser Ala Val Arg His Val Val Gly Gln Lys Pro Thr Ser Asp Arg Pro 
        515                 520                 525             

Tyr Ile Trp Asn Asp Asn Glu Gln Gly Leu Asp Glu His Ile Ala Arg 
    530                 535                 540                 

Ile Ser Ala Asp Ile Ala Ala Gly Gly Val Ile Val Gln Ala Val Gln 
545                 550                 555                 560 

Asp Ile Leu Pro Cys Leu His 
                565         

<210>  5
<211>  43
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create cysteine to serine 
       substitution at position 318 (forward)

<400>  5
caagatcgtt actcactccg atcccttccc cagtatttgg ggc                       43

<210>  6
<211>  43
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create cysteine to serine 
       substitution at position 318 (reverse)

<400>  6
gccccaaata ctggggaagg gatcggagtg agtaacgatc ttg                       43

<210>  7
<211>  567
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Cysteine to serine substitution at position 64 in Anabaena 
       variabilis PAL

<400>  7

Met Lys Thr Leu Ser Gln Ala Gln Ser Lys Thr Ser Ser Gln Gln Phe 
1               5                   10                  15      

Ser Phe Thr Gly Asn Ser Ser Ala Asn Val Ile Ile Gly Asn Gln Lys 
            20                  25                  30          

Leu Thr Ile Asn Asp Val Ala Arg Val Ala Arg Asn Gly Thr Leu Val 
        35                  40                  45              

Ser Leu Thr Asn Asn Thr Asp Ile Leu Gln Gly Ile Gln Ala Ser Ser 
    50                  55                  60                  

Asp Tyr Ile Asn Asn Ala Val Glu Ser Gly Glu Pro Ile Tyr Gly Val 
65                  70                  75                  80  

Thr Ser Gly Phe Gly Gly Met Ala Asn Val Ala Ile Ser Arg Glu Gln 
                85                  90                  95      

Ala Ser Glu Leu Gln Thr Asn Leu Val Trp Phe Leu Lys Thr Gly Ala 
            100                 105                 110         

Gly Asn Lys Leu Pro Leu Ala Asp Val Arg Ala Ala Met Leu Leu Arg 
        115                 120                 125             

Ala Asn Ser His Met Arg Gly Ala Ser Gly Ile Arg Leu Glu Leu Ile 
    130                 135                 140                 

Lys Arg Met Glu Ile Phe Leu Asn Ala Gly Val Thr Pro Tyr Val Tyr 
145                 150                 155                 160 

Glu Phe Gly Ser Ile Gly Ala Ser Gly Asp Leu Val Pro Leu Ser Tyr 
                165                 170                 175     

Ile Thr Gly Ser Leu Ile Gly Leu Asp Pro Ser Phe Lys Val Asp Phe 
            180                 185                 190         

Asn Gly Lys Glu Met Asp Ala Pro Thr Ala Leu Arg Gln Leu Asn Leu 
        195                 200                 205             

Ser Pro Leu Thr Leu Leu Pro Lys Glu Gly Leu Ala Met Met Asn Gly 
    210                 215                 220                 

Thr Ser Val Met Thr Gly Ile Ala Ala Asn Cys Val Tyr Asp Thr Gln 
225                 230                 235                 240 

Ile Leu Thr Ala Ile Ala Met Gly Val His Ala Leu Asp Ile Gln Ala 
                245                 250                 255     

Leu Asn Gly Thr Asn Gln Ser Phe His Pro Phe Ile His Asn Ser Lys 
            260                 265                 270         

Pro His Pro Gly Gln Leu Trp Ala Ala Asp Gln Met Ile Ser Leu Leu 
        275                 280                 285             

Ala Asn Ser Gln Leu Val Arg Asp Glu Leu Asp Gly Lys His Asp Tyr 
    290                 295                 300                 

Arg Asp His Glu Leu Ile Gln Asp Arg Tyr Ser Leu Arg Cys Leu Pro 
305                 310                 315                 320 

Gln Tyr Leu Gly Pro Ile Val Asp Gly Ile Ser Gln Ile Ala Lys Gln 
                325                 330                 335     

Ile Glu Ile Glu Ile Asn Ser Val Thr Asp Asn Pro Leu Ile Asp Val 
            340                 345                 350         

Asp Asn Gln Ala Ser Tyr His Gly Gly Asn Phe Leu Gly Gln Tyr Val 
        355                 360                 365             

Gly Met Gly Met Asp His Leu Arg Tyr Tyr Ile Gly Leu Leu Ala Lys 
    370                 375                 380                 

His Leu Asp Val Gln Ile Ala Leu Leu Ala Ser Pro Glu Phe Ser Asn 
385                 390                 395                 400 

Gly Leu Pro Pro Ser Leu Leu Gly Asn Arg Glu Arg Lys Val Asn Met 
                405                 410                 415     

Gly Leu Lys Gly Leu Gln Ile Cys Gly Asn Ser Ile Met Pro Leu Leu 
            420                 425                 430         

Thr Phe Tyr Gly Asn Ser Ile Ala Asp Arg Phe Pro Thr His Ala Glu 
        435                 440                 445             

Gln Phe Asn Gln Asn Ile Asn Ser Gln Gly Tyr Thr Ser Ala Thr Leu 
    450                 455                 460                 

Ala Arg Arg Ser Val Asp Ile Phe Gln Asn Tyr Val Ala Ile Ala Leu 
465                 470                 475                 480 

Met Phe Gly Val Gln Ala Val Asp Leu Arg Thr Tyr Lys Lys Thr Gly 
                485                 490                 495     

His Tyr Asp Ala Arg Ala Cys Leu Ser Pro Ala Thr Glu Arg Leu Tyr 
            500                 505                 510         

Ser Ala Val Arg His Val Val Gly Gln Lys Pro Thr Ser Asp Arg Pro 
        515                 520                 525             

Tyr Ile Trp Asn Asp Asn Glu Gln Gly Leu Asp Glu His Ile Ala Arg 
    530                 535                 540                 

Ile Ser Ala Asp Ile Ala Ala Gly Gly Val Ile Val Gln Ala Val Gln 
545                 550                 555                 560 

Asp Ile Leu Pro Cys Leu His 
                565         

<210>  8
<211>  567
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Cysteine to serine substitution at position 318 in Anabaena 
       variabilis PAL

<400>  8

Met Lys Thr Leu Ser Gln Ala Gln Ser Lys Thr Ser Ser Gln Gln Phe 
1               5                   10                  15      

Ser Phe Thr Gly Asn Ser Ser Ala Asn Val Ile Ile Gly Asn Gln Lys 
            20                  25                  30          

Leu Thr Ile Asn Asp Val Ala Arg Val Ala Arg Asn Gly Thr Leu Val 
        35                  40                  45              

Ser Leu Thr Asn Asn Thr Asp Ile Leu Gln Gly Ile Gln Ala Ser Cys 
    50                  55                  60                  

Asp Tyr Ile Asn Asn Ala Val Glu Ser Gly Glu Pro Ile Tyr Gly Val 
65                  70                  75                  80  

Thr Ser Gly Phe Gly Gly Met Ala Asn Val Ala Ile Ser Arg Glu Gln 
                85                  90                  95      

Ala Ser Glu Leu Gln Thr Asn Leu Val Trp Phe Leu Lys Thr Gly Ala 
            100                 105                 110         

Gly Asn Lys Leu Pro Leu Ala Asp Val Arg Ala Ala Met Leu Leu Arg 
        115                 120                 125             

Ala Asn Ser His Met Arg Gly Ala Ser Gly Ile Arg Leu Glu Leu Ile 
    130                 135                 140                 

Lys Arg Met Glu Ile Phe Leu Asn Ala Gly Val Thr Pro Tyr Val Tyr 
145                 150                 155                 160 

Glu Phe Gly Ser Ile Gly Ala Ser Gly Asp Leu Val Pro Leu Ser Tyr 
                165                 170                 175     

Ile Thr Gly Ser Leu Ile Gly Leu Asp Pro Ser Phe Lys Val Asp Phe 
            180                 185                 190         

Asn Gly Lys Glu Met Asp Ala Pro Thr Ala Leu Arg Gln Leu Asn Leu 
        195                 200                 205             

Ser Pro Leu Thr Leu Leu Pro Lys Glu Gly Leu Ala Met Met Asn Gly 
    210                 215                 220                 

Thr Ser Val Met Thr Gly Ile Ala Ala Asn Cys Val Tyr Asp Thr Gln 
225                 230                 235                 240 

Ile Leu Thr Ala Ile Ala Met Gly Val His Ala Leu Asp Ile Gln Ala 
                245                 250                 255     

Leu Asn Gly Thr Asn Gln Ser Phe His Pro Phe Ile His Asn Ser Lys 
            260                 265                 270         

Pro His Pro Gly Gln Leu Trp Ala Ala Asp Gln Met Ile Ser Leu Leu 
        275                 280                 285             

Ala Asn Ser Gln Leu Val Arg Asp Glu Leu Asp Gly Lys His Asp Tyr 
    290                 295                 300                 

Arg Asp His Glu Leu Ile Gln Asp Arg Tyr Ser Leu Arg Ser Leu Pro 
305                 310                 315                 320 

Gln Tyr Leu Gly Pro Ile Val Asp Gly Ile Ser Gln Ile Ala Lys Gln 
                325                 330                 335     

Ile Glu Ile Glu Ile Asn Ser Val Thr Asp Asn Pro Leu Ile Asp Val 
            340                 345                 350         

Asp Asn Gln Ala Ser Tyr His Gly Gly Asn Phe Leu Gly Gln Tyr Val 
        355                 360                 365             

Gly Met Gly Met Asp His Leu Arg Tyr Tyr Ile Gly Leu Leu Ala Lys 
    370                 375                 380                 

His Leu Asp Val Gln Ile Ala Leu Leu Ala Ser Pro Glu Phe Ser Asn 
385                 390                 395                 400 

Gly Leu Pro Pro Ser Leu Leu Gly Asn Arg Glu Arg Lys Val Asn Met 
                405                 410                 415     

Gly Leu Lys Gly Leu Gln Ile Cys Gly Asn Ser Ile Met Pro Leu Leu 
            420                 425                 430         

Thr Phe Tyr Gly Asn Ser Ile Ala Asp Arg Phe Pro Thr His Ala Glu 
        435                 440                 445             

Gln Phe Asn Gln Asn Ile Asn Ser Gln Gly Tyr Thr Ser Ala Thr Leu 
    450                 455                 460                 

Ala Arg Arg Ser Val Asp Ile Phe Gln Asn Tyr Val Ala Ile Ala Leu 
465                 470                 475                 480 

Met Phe Gly Val Gln Ala Val Asp Leu Arg Thr Tyr Lys Lys Thr Gly 
                485                 490                 495     

His Tyr Asp Ala Arg Ala Cys Leu Ser Pro Ala Thr Glu Arg Leu Tyr 
            500                 505                 510         

Ser Ala Val Arg His Val Val Gly Gln Lys Pro Thr Ser Asp Arg Pro 
        515                 520                 525             

Tyr Ile Trp Asn Asp Asn Glu Gln Gly Leu Asp Glu His Ile Ala Arg 
    530                 535                 540                 

Ile Ser Ala Asp Ile Ala Ala Gly Gly Val Ile Val Gln Ala Val Gln 
545                 550                 555                 560 

Asp Ile Leu Pro Cys Leu His 
                565         

<210>  9
<211>  567
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Cysteine to serine substitution at position 503 in Anabaena 
       variabilis PAL

<400>  9

Met Lys Thr Leu Ser Gln Ala Gln Ser Lys Thr Ser Ser Gln Gln Phe 
1               5                   10                  15      

Ser Phe Thr Gly Asn Ser Ser Ala Asn Val Ile Ile Gly Asn Gln Lys 
            20                  25                  30          

Leu Thr Ile Asn Asp Val Ala Arg Val Ala Arg Asn Gly Thr Leu Val 
        35                  40                  45              

Ser Leu Thr Asn Asn Thr Asp Ile Leu Gln Gly Ile Gln Ala Ser Cys 
    50                  55                  60                  

Asp Tyr Ile Asn Asn Ala Val Glu Ser Gly Glu Pro Ile Tyr Gly Val 
65                  70                  75                  80  

Thr Ser Gly Phe Gly Gly Met Ala Asn Val Ala Ile Ser Arg Glu Gln 
                85                  90                  95      

Ala Ser Glu Leu Gln Thr Asn Leu Val Trp Phe Leu Lys Thr Gly Ala 
            100                 105                 110         

Gly Asn Lys Leu Pro Leu Ala Asp Val Arg Ala Ala Met Leu Leu Arg 
        115                 120                 125             

Ala Asn Ser His Met Arg Gly Ala Ser Gly Ile Arg Leu Glu Leu Ile 
    130                 135                 140                 

Lys Arg Met Glu Ile Phe Leu Asn Ala Gly Val Thr Pro Tyr Val Tyr 
145                 150                 155                 160 

Glu Phe Gly Ser Ile Gly Ala Ser Gly Asp Leu Val Pro Leu Ser Tyr 
                165                 170                 175     

Ile Thr Gly Ser Leu Ile Gly Leu Asp Pro Ser Phe Lys Val Asp Phe 
            180                 185                 190         

Asn Gly Lys Glu Met Asp Ala Pro Thr Ala Leu Arg Gln Leu Asn Leu 
        195                 200                 205             

Ser Pro Leu Thr Leu Leu Pro Lys Glu Gly Leu Ala Met Met Asn Gly 
    210                 215                 220                 

Thr Ser Val Met Thr Gly Ile Ala Ala Asn Cys Val Tyr Asp Thr Gln 
225                 230                 235                 240 

Ile Leu Thr Ala Ile Ala Met Gly Val His Ala Leu Asp Ile Gln Ala 
                245                 250                 255     

Leu Asn Gly Thr Asn Gln Ser Phe His Pro Phe Ile His Asn Ser Lys 
            260                 265                 270         

Pro His Pro Gly Gln Leu Trp Ala Ala Asp Gln Met Ile Ser Leu Leu 
        275                 280                 285             

Ala Asn Ser Gln Leu Val Arg Asp Glu Leu Asp Gly Lys His Asp Tyr 
    290                 295                 300                 

Arg Asp His Glu Leu Ile Gln Asp Arg Tyr Ser Leu Arg Cys Leu Pro 
305                 310                 315                 320 

Gln Tyr Leu Gly Pro Ile Val Asp Gly Ile Ser Gln Ile Ala Lys Gln 
                325                 330                 335     

Ile Glu Ile Glu Ile Asn Ser Val Thr Asp Asn Pro Leu Ile Asp Val 
            340                 345                 350         

Asp Asn Gln Ala Ser Tyr His Gly Gly Asn Phe Leu Gly Gln Tyr Val 
        355                 360                 365             

Gly Met Gly Met Asp His Leu Arg Tyr Tyr Ile Gly Leu Leu Ala Lys 
    370                 375                 380                 

His Leu Asp Val Gln Ile Ala Leu Leu Ala Ser Pro Glu Phe Ser Asn 
385                 390                 395                 400 

Gly Leu Pro Pro Ser Leu Leu Gly Asn Arg Glu Arg Lys Val Asn Met 
                405                 410                 415     

Gly Leu Lys Gly Leu Gln Ile Cys Gly Asn Ser Ile Met Pro Leu Leu 
            420                 425                 430         

Thr Phe Tyr Gly Asn Ser Ile Ala Asp Arg Phe Pro Thr His Ala Glu 
        435                 440                 445             

Gln Phe Asn Gln Asn Ile Asn Ser Gln Gly Tyr Thr Ser Ala Thr Leu 
    450                 455                 460                 

Ala Arg Arg Ser Val Asp Ile Phe Gln Asn Tyr Val Ala Ile Ala Leu 
465                 470                 475                 480 

Met Phe Gly Val Gln Ala Val Asp Leu Arg Thr Tyr Lys Lys Thr Gly 
                485                 490                 495     

His Tyr Asp Ala Arg Ala Ser Leu Ser Pro Ala Thr Glu Arg Leu Tyr 
            500                 505                 510         

Ser Ala Val Arg His Val Val Gly Gln Lys Pro Thr Ser Asp Arg Pro 
        515                 520                 525             

Tyr Ile Trp Asn Asp Asn Glu Gln Gly Leu Asp Glu His Ile Ala Arg 
    530                 535                 540                 

Ile Ser Ala Asp Ile Ala Ala Gly Gly Val Ile Val Gln Ala Val Gln 
545                 550                 555                 560 

Asp Ile Leu Pro Cys Leu His 
                565         

<210>  10
<211>  567
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Cysteine to serine substition at position 565 in Anabaena 
       variabilis PAL

<400>  10

Met Lys Thr Leu Ser Gln Ala Gln Ser Lys Thr Ser Ser Gln Gln Phe 
1               5                   10                  15      

Ser Phe Thr Gly Asn Ser Ser Ala Asn Val Ile Ile Gly Asn Gln Lys 
            20                  25                  30          

Leu Thr Ile Asn Asp Val Ala Arg Val Ala Arg Asn Gly Thr Leu Val 
        35                  40                  45              

Ser Leu Thr Asn Asn Thr Asp Ile Leu Gln Gly Ile Gln Ala Ser Cys 
    50                  55                  60                  

Asp Tyr Ile Asn Asn Ala Val Glu Ser Gly Glu Pro Ile Tyr Gly Val 
65                  70                  75                  80  

Thr Ser Gly Phe Gly Gly Met Ala Asn Val Ala Ile Ser Arg Glu Gln 
                85                  90                  95      

Ala Ser Glu Leu Gln Thr Asn Leu Val Trp Phe Leu Lys Thr Gly Ala 
            100                 105                 110         

Gly Asn Lys Leu Pro Leu Ala Asp Val Arg Ala Ala Met Leu Leu Arg 
        115                 120                 125             

Ala Asn Ser His Met Arg Gly Ala Ser Gly Ile Arg Leu Glu Leu Ile 
    130                 135                 140                 

Lys Arg Met Glu Ile Phe Leu Asn Ala Gly Val Thr Pro Tyr Val Tyr 
145                 150                 155                 160 

Glu Phe Gly Ser Ile Gly Ala Ser Gly Asp Leu Val Pro Leu Ser Tyr 
                165                 170                 175     

Ile Thr Gly Ser Leu Ile Gly Leu Asp Pro Ser Phe Lys Val Asp Phe 
            180                 185                 190         

Asn Gly Lys Glu Met Asp Ala Pro Thr Ala Leu Arg Gln Leu Asn Leu 
        195                 200                 205             

Ser Pro Leu Thr Leu Leu Pro Lys Glu Gly Leu Ala Met Met Asn Gly 
    210                 215                 220                 

Thr Ser Val Met Thr Gly Ile Ala Ala Asn Cys Val Tyr Asp Thr Gln 
225                 230                 235                 240 

Ile Leu Thr Ala Ile Ala Met Gly Val His Ala Leu Asp Ile Gln Ala 
                245                 250                 255     

Leu Asn Gly Thr Asn Gln Ser Phe His Pro Phe Ile His Asn Ser Lys 
            260                 265                 270         

Pro His Pro Gly Gln Leu Trp Ala Ala Asp Gln Met Ile Ser Leu Leu 
        275                 280                 285             

Ala Asn Ser Gln Leu Val Arg Asp Glu Leu Asp Gly Lys His Asp Tyr 
    290                 295                 300                 

Arg Asp His Glu Leu Ile Gln Asp Arg Tyr Ser Leu Arg Cys Leu Pro 
305                 310                 315                 320 

Gln Tyr Leu Gly Pro Ile Val Asp Gly Ile Ser Gln Ile Ala Lys Gln 
                325                 330                 335     

Ile Glu Ile Glu Ile Asn Ser Val Thr Asp Asn Pro Leu Ile Asp Val 
            340                 345                 350         

Asp Asn Gln Ala Ser Tyr His Gly Gly Asn Phe Leu Gly Gln Tyr Val 
        355                 360                 365             

Gly Met Gly Met Asp His Leu Arg Tyr Tyr Ile Gly Leu Leu Ala Lys 
    370                 375                 380                 

His Leu Asp Val Gln Ile Ala Leu Leu Ala Ser Pro Glu Phe Ser Asn 
385                 390                 395                 400 

Gly Leu Pro Pro Ser Leu Leu Gly Asn Arg Glu Arg Lys Val Asn Met 
                405                 410                 415     

Gly Leu Lys Gly Leu Gln Ile Cys Gly Asn Ser Ile Met Pro Leu Leu 
            420                 425                 430         

Thr Phe Tyr Gly Asn Ser Ile Ala Asp Arg Phe Pro Thr His Ala Glu 
        435                 440                 445             

Gln Phe Asn Gln Asn Ile Asn Ser Gln Gly Tyr Thr Ser Ala Thr Leu 
    450                 455                 460                 

Ala Arg Arg Ser Val Asp Ile Phe Gln Asn Tyr Val Ala Ile Ala Leu 
465                 470                 475                 480 

Met Phe Gly Val Gln Ala Val Asp Leu Arg Thr Tyr Lys Lys Thr Gly 
                485                 490                 495     

His Tyr Asp Ala Arg Ala Cys Leu Ser Pro Ala Thr Glu Arg Leu Tyr 
            500                 505                 510         

Ser Ala Val Arg His Val Val Gly Gln Lys Pro Thr Ser Asp Arg Pro 
        515                 520                 525             

Tyr Ile Trp Asn Asp Asn Glu Gln Gly Leu Asp Glu His Ile Ala Arg 
    530                 535                 540                 

Ile Ser Ala Asp Ile Ala Ala Gly Gly Val Ile Val Gln Ala Val Gln 
545                 550                 555                 560 

Asp Ile Leu Pro Ser Leu His 
                565         

<210>  11
<211>  567
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Cysteine to serine substitutions at positions 565 and 503 in 
       Anabaena variabilis PAL

<400>  11

Met Lys Thr Leu Ser Gln Ala Gln Ser Lys Thr Ser Ser Gln Gln Phe 
1               5                   10                  15      

Ser Phe Thr Gly Asn Ser Ser Ala Asn Val Ile Ile Gly Asn Gln Lys 
            20                  25                  30          

Leu Thr Ile Asn Asp Val Ala Arg Val Ala Arg Asn Gly Thr Leu Val 
        35                  40                  45              

Ser Leu Thr Asn Asn Thr Asp Ile Leu Gln Gly Ile Gln Ala Ser Cys 
    50                  55                  60                  

Asp Tyr Ile Asn Asn Ala Val Glu Ser Gly Glu Pro Ile Tyr Gly Val 
65                  70                  75                  80  

Thr Ser Gly Phe Gly Gly Met Ala Asn Val Ala Ile Ser Arg Glu Gln 
                85                  90                  95      

Ala Ser Glu Leu Gln Thr Asn Leu Val Trp Phe Leu Lys Thr Gly Ala 
            100                 105                 110         

Gly Asn Lys Leu Pro Leu Ala Asp Val Arg Ala Ala Met Leu Leu Arg 
        115                 120                 125             

Ala Asn Ser His Met Arg Gly Ala Ser Gly Ile Arg Leu Glu Leu Ile 
    130                 135                 140                 

Lys Arg Met Glu Ile Phe Leu Asn Ala Gly Val Thr Pro Tyr Val Tyr 
145                 150                 155                 160 

Glu Phe Gly Ser Ile Gly Ala Ser Gly Asp Leu Val Pro Leu Ser Tyr 
                165                 170                 175     

Ile Thr Gly Ser Leu Ile Gly Leu Asp Pro Ser Phe Lys Val Asp Phe 
            180                 185                 190         

Asn Gly Lys Glu Met Asp Ala Pro Thr Ala Leu Arg Gln Leu Asn Leu 
        195                 200                 205             

Ser Pro Leu Thr Leu Leu Pro Lys Glu Gly Leu Ala Met Met Asn Gly 
    210                 215                 220                 

Thr Ser Val Met Thr Gly Ile Ala Ala Asn Cys Val Tyr Asp Thr Gln 
225                 230                 235                 240 

Ile Leu Thr Ala Ile Ala Met Gly Val His Ala Leu Asp Ile Gln Ala 
                245                 250                 255     

Leu Asn Gly Thr Asn Gln Ser Phe His Pro Phe Ile His Asn Ser Lys 
            260                 265                 270         

Pro His Pro Gly Gln Leu Trp Ala Ala Asp Gln Met Ile Ser Leu Leu 
        275                 280                 285             

Ala Asn Ser Gln Leu Val Arg Asp Glu Leu Asp Gly Lys His Asp Tyr 
    290                 295                 300                 

Arg Asp His Glu Leu Ile Gln Asp Arg Tyr Ser Leu Arg Cys Leu Pro 
305                 310                 315                 320 

Gln Tyr Leu Gly Pro Ile Val Asp Gly Ile Ser Gln Ile Ala Lys Gln 
                325                 330                 335     

Ile Glu Ile Glu Ile Asn Ser Val Thr Asp Asn Pro Leu Ile Asp Val 
            340                 345                 350         

Asp Asn Gln Ala Ser Tyr His Gly Gly Asn Phe Leu Gly Gln Tyr Val 
        355                 360                 365             

Gly Met Gly Met Asp His Leu Arg Tyr Tyr Ile Gly Leu Leu Ala Lys 
    370                 375                 380                 

His Leu Asp Val Gln Ile Ala Leu Leu Ala Ser Pro Glu Phe Ser Asn 
385                 390                 395                 400 

Gly Leu Pro Pro Ser Leu Leu Gly Asn Arg Glu Arg Lys Val Asn Met 
                405                 410                 415     

Gly Leu Lys Gly Leu Gln Ile Cys Gly Asn Ser Ile Met Pro Leu Leu 
            420                 425                 430         

Thr Phe Tyr Gly Asn Ser Ile Ala Asp Arg Phe Pro Thr His Ala Glu 
        435                 440                 445             

Gln Phe Asn Gln Asn Ile Asn Ser Gln Gly Tyr Thr Ser Ala Thr Leu 
    450                 455                 460                 

Ala Arg Arg Ser Val Asp Ile Phe Gln Asn Tyr Val Ala Ile Ala Leu 
465                 470                 475                 480 

Met Phe Gly Val Gln Ala Val Asp Leu Arg Thr Tyr Lys Lys Thr Gly 
                485                 490                 495     

His Tyr Asp Ala Arg Ala Ser Leu Ser Pro Ala Thr Glu Arg Leu Tyr 
            500                 505                 510         

Ser Ala Val Arg His Val Val Gly Gln Lys Pro Thr Ser Asp Arg Pro 
        515                 520                 525             

Tyr Ile Trp Asn Asp Asn Glu Gln Gly Leu Asp Glu His Ile Ala Arg 
    530                 535                 540                 

Ile Ser Ala Asp Ile Ala Ala Gly Gly Val Ile Val Gln Ala Val Gln 
545                 550                 555                 560 

Asp Ile Leu Pro Ser Leu His 
                565         

<210>  12
<211>  38
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nostoc punctiforme PAL primer 1 (forward)

<400>  12
cactgtcata tgaatataac atctctacaa cagaacat                             38

<210>  13
<211>  45
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nostoc punctiforme PAL primer 2 (reverse)

<400>  13
gacagtggcg gccgctcacg ttgactttaa gctcgaaaaa atatg                     45

<210>  14
<211>  38
<212>  DNA
<213>  Artificial Sequence 

<220>
<223>  Anabaena variabilis PAL primer 1 (forward, N-terminal fragment)

<400>  14
cactgtgcta gcatgaagac actatctcaa gcacaaag                             38

<210>  15
<211>  49
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer 2 (reverse, N-terminal fragment)

<400>  15
ggaaatttcc tccatgatag ctggcttggt tatcaacatc aattagtgg                 49

<210>  16
<211>  49
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer 3 (forward, C-terminal fragment)

<400>  16
ccactaattg atgttgataa ccaagccagc tatcatggag gaaatttcc                 49

<210>  17
<211>  41
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer 4 (reverse, C-terminal fragment)

<400>  17
cactgtgcgg ccgcttaatg caagcagggt aagatatctt g                         41

<210>  18
<211>  35
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL forward primer

<400>  18
cactgtcata tgaagacact atctcaagca caaag                                35

<210>  19
<211>  36
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL reverse primer

<400>  19
cactgtctcg agatgcaagc agggtaagat atcttg                               36

<210>  20
<211>  38
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create 5' NheI site and delete 
       internal NheI site (forward, N-terminal)

<400>  20
cactgtgcta gcatgaagac actatctcaa gcacaaag                             38

<210>  21
<211>  49
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create 5' NheI site and delete 
       internal NheI site  (reverse, N-terminal )

<400>  21
ggaaatttcc tccatgatag ctggcttggt tatcaacatc aattagtgg                 49

<210>  22
<211>  49
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create 5' NheI site and delete 
       internal NheI site (forward, C-terminal fragment)

<400>  22
ccactaattg atgttgataa ccaagccagc tatcatggag gaaatttcc                 49

<210>  23
<211>  41
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create 5' NheI site and delete 
       internal NheI site  (reverse, C-terminal fragment)

<400>  23
acagtggcgg ccgcttaatg caagcagggt aagatatctt g                         41

<210>  24
<211>  38
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create 5' NheI site and 3' SmaI
       site (forward)

<400>  24
cactgtgaat tcatgaagac actatctcaa gcacaaag                             38

<210>  25
<211>  37
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create 5' NheI site and 3' SmaI
       site (reverse)

<400>  25
cactgtcccg ggttaatgca agcagggtaa gatatct                              37

<210>  26
<211>  41
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create cysteine to serine 
       substitution at position 503 (forward)

<400>  26
gtcattacga tgcacgcgcc tctctatcac ctgcaactga g                         41

<210>  27
<211>  41
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create cysteine to serine 
       substitution at position 503 (reverse)

<400>  27
ctcagttgca ggtgatagag aggcgcgtgc atcgtaatga c                         41

<210>  28
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create cysteine to serine 
       substitution at position 565 (forward)

<400>  28
cagttcaaga tatcttaccc tccttgcatt aacccgggct gc                        42

<210>  29
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create cysteine to serine 
       substitution at position 565 (reverse)

<400>  29
gcagcccggg ttaatgcaag gagggtaaga tatcttgaac tg                        42

<210>  30
<211>  44
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create cysteine to serine 
       substitution at position 64 (forward)

<400>  30
gcagggtatt caggcatctt ctgattacat taataatgct gttg                      44

<210>  31
<211>  44
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Anabaena variabilis PAL primer to create cysteine to serine 
       substitution at position 64 (reverse)

<400>  31
caacagcatt attaatgtaa tcagaagatg cctgaatacc ctgc                      44

