                         SEQUENCE LISTING

<110>  Montana State University
       Pascual, David
       Maddaloni, Massimo
 
<120>  Recombinant Lactococcus lactis Expressing Escherichia coli 
       Colonization Factor Antigen I (CFA/I) Fimbriae and Their Methods 
       of Use

<130>  MONT-144/01US

<150>  61/704,672
<151>  2012-09-24

<160>  12    

<170>  PatentIn version 3.5

<210>  1
<211>  5565
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Engineered CFA/I Operon

<400>  1
tctagaagtc ttataactat actgacaata gaaacattaa caaatctaaa acagtcttaa       60

ttctatcttg agaaagtatt ggtaataata ttattgtcga taacgcgagc ataataaacg      120

gctctgatta aattctgaag tttgttagat acaatgattt cgttcgaagg aactacaaaa      180

taaattattc tagaccaggt gatatcaata tgcgaaaaga actatgaata tccactccat      240

ttttggttgc catttgttaa cgctgcctcc tctccctagt gctataataa aacaggccca      300

ttttggaaca gacttctact attttgttgt agatctgggc cccatttggc agtttattct      360

tgacatgtag tgagggggct ggtataatca catagtactg tttgattctt cagcaagact      420

ggtacctcat gagagttata gactcatgga tcttgctttg aagggttttg tacattatag      480

gctcctatca catgctgaac ctatggccta ttacattttt ttatatttca aggaggaccg      540

gtaccaggta ctagtgtcgc gaggagggct agttctagaa tgaaaaaaaa gattatctca      600

gctattttaa tgtctacagt gatactttct gctgcagccc cgttgtcagg tgtttacgct      660

gctagcgaga aaaatattac tgtaacagct agtgttgatc ctgcaattga tcttttgcaa      720

gctgatggca atgctctgcc atcagctgta aagttagctt attctcccgc atcaaaaact      780

tttgaaagtt acagagtaat gactcaagta catacaaacg atgcaactaa aaaagtaatt      840

gttaaacttg ctgatacacc acagcttaca gatgttctga attcaactgt tcaaatgcct      900

atcagtgtgt catggggagg acaagtatta tctacaacag ccaaagaatt tgaagctgct      960

gctttgggat attctgcatc cggtgtaaat ggcgtatcat cttctcaaga gttagtaatt     1020

agcgctgcac ctaaaactgc cggtaccgcc ccaactgcag gaaactattc aggagtagta     1080

tctcttgtaa tgactttggg atcctgaggg cccaggaggg tttaaacatg aaaaagatta     1140

atctggcatt attgaccctc gcaactttga tgggtgtttc ttcaacagct gttgtatttg     1200

ccggcaactt tatgatatat ccaatatcaa aagatttaaa gaatggaaat agcgagttag     1260

ttcgtgttta ttcaaaatca aaagagatac aatatataaa aatatataca aagaagatta     1320

ttaatcccgg tacaactgaa gaatataagg ttgatatacc caattgggat ggtgggcttg     1380

tagtcactcc tcagaaagtt atcctccctg caggagcgag taaatcaata cgtttaactc     1440

aatttaaaat accaaaaaaa gaggaagttt acagagtata ttttgaggcg gtaaaaccag     1500

atagcaaaga aaatgtaatt gataataaaa aactaacaac agagctatct gttaatataa     1560

tttatgcggc tctaattaga tctttaccaa gtgaacaaaa tatatcacta aatatttcta     1620

gaaatgcaaa aaaaaatata attatttata ataatgggaa tgttagagca ggcgttaaag     1680

atatttattt ttgtaagtca tctaatatcg atgataactg tgtaaaaaaa gcgtataaca     1740

agaatatata tccagaaaaa gtcatttgag ccggctcgcg aatgaaaaag attttgatca     1800

ctacgacatt agcacttgct ctcctgtctt taggtgcagc tagcgttacc ggagatatac     1860

ccgactcttt ccgtgattta tggggagaac aagatgaatt ttatgaagta aaactatatg     1920

ggcaaactct aggaatacat cgaattaaaa caaccccaac acatattaag ttttattcac     1980

ccgaaagcat tttagataaa ataaatttaa aaaaagaaaa ggaaaaggaa ttgagtgttt     2040

tttttactaa ttctttttca agaaatggca atatgagttg tcagggtaac actactatac     2100

agtataactg caattacatt aaaacaaaat cagtagatgt catcgttgat gatgttgata     2160

atgttgttaa cctttttata ggtaatgaat ttctggattc tgaagcacac aatgatgaat     2220

atcatcaatt atcacggaat gtaaaaaaag cttttataca aagccagaca attaatctct     2280

cagattctgg aaagtataaa agattgtcta tttcagggaa tagcgcgctg ggtattacag     2340

atacaagtta tgctgtctta aattggtgga tgaattacaa taaatctaat ggttacagca     2400

acaacgaaaa aacaatcaat agtttatact ttagacatga tttagataag agatattatt     2460

atcaatttgg acgaatggat cgtacagatt tatcacaaag tattagcggg agctttaatt     2520

ttaacttact tcctttaccc gatattgatg gtatacggac aggaaccaca caatcttata     2580

tcaaaaatac agataagttt atcgcatccc ctgtaactgt tatgttaact aatttttcca     2640

gagtggaagc ttttcgcaat gatcaattat tgggcgtatg gtatttagat tctggagtaa     2700

atgaattaga tacagctcgt ttaccttatg gcagttacga tcttaaatta aaaatttttg     2760

aaaacactca attagttcgt gaagaaataa ttccttttaa taaaggaaga agctctattg     2820

gtgatatgca atgggatatt ttcgttcagg gagggaatat tgttaatgat aatgatcgtt     2880

acatagaaaa acaaaataat cataagtcat cgattaatac tgggctacgt ttaccaatta     2940

cgaaaaatat ctctgttcaa cagggagtat ctgttataga taataaaagt tattatgaag     3000

gaagtctgaa atggaattcc ggcattctat ctggctcact aaatagtgag ttcagttttc     3060

tttggggaga taatgcaaaa ggtaattatc aaagtatctc gtataccgat ggatttagct     3120

tatcatttta tcataatgat aagcgggtcg ataattgtgg aagaaattac aatgctggtt     3180

ggagtggatg ctacgaatca tattcggcat ctttaagtat tcctttatta ggatggacaa     3240

gtactctggg atatagtgac acttatagtg aatcagtgta taaaagccat attctttctg     3300

aatatggctt ttataatcaa aacatatata aagggagaac ccaaagatgg caactgactt     3360

catccacctc tttaaaatgg atggattata attttatgcc agcaattgga atatataaca     3420

gtgaacaaag acaactgact gataaaggcg gatatatatc tgtaactatc acccgagcca     3480

gcagagaaaa ttcattaaat acagggtatt cttacaacta ttccagagga aactattctt     3540

ctaacgaatt atttgttgat ggatatatga catcaacaaa taatggtgat tatcatgagg     3600

caggaatgcg ttttaataaa aatagacata atgcagaagg tagactttca ggtcgtataa     3660

acaatcgatt tggagattta aatggttcat tcagcatgaa taaaaacaga aacaccaaca     3720

gtaccaatca ttctctcact ggtggttata attcctcatt tgctcttaca agtgatggat     3780

tttactgggg aggaagtaca gctggtttga caaaactggc tggcggtatt atcaaggtta     3840

aatcaaacga tactaaaaaa aacttggtaa aagtgactgg gacattgtac ggtgattatt     3900

cgctagggag caacgataat gcttttattc ctgtaccagc attaactcca gccagtttaa     3960

tcattgaaga taataattat ggtgataata atatttctat acttgcgcca acaaacaacg     4020

atatgtttat gttgccgggt aatgtttatc ctgttgaaat tgaaaccaaa gtaagtgttt     4080

cttatattgg tagaggtttt gacccaaacg gcacgccact ttctggcgca catgttttga     4140

atgaaccaca tgttatcctg gatgaggacg gtggattttc gtttgaatat acaggtaatg     4200

agaaaacact ttttttatta aagggcagga ctatttatac atgtcaactg gggaaaaata     4260

aagttcataa aggcattgtt ttcgtcgggg atgttatatg tgatattaat agcacaagtt     4320

ccttaccaga tgaatttgta aagaacccac gtgtgcagga tttgctggca aagaatgata     4380

aaggataaac ggctagcgtt taaaccgact cagtggtcga catgcaaagg aaaaagaaag     4440

ggctatcgtt cttgttagcc ggtacagtcg ctttaggggc gctggctgtc ttgccagtcg     4500

gcgaaatcca agcaaaggcg gccgcagata aaaatcccgg aagtgaaaac atgactaata     4560

ctattggtcc ccatgacagg gggggatctt cccccatata taatatctta aattcctatc     4620

ttacagcata caatggaagc catcatctgt atgataggat gagtttttta tgtttgtctt     4680

ctcaaaatac actgaatgga gcatgcccaa gcagtgatgc ccctggcact gctacaattg     4740

atggcgaaac aaatataaca ttacaattta cggaaaaaag aagtctaatt aaaagagaac     4800

tgcaaattaa aggctataaa caatttttgt tcaaaaatgc taattgccca tctaaactag     4860

cacttaactc atctcatttt caatgtaata gagaacaagc ttcaggtgct actttatcgt     4920

tatacatacc agctggtgaa ttaaataaat taccttttgg gggggtctgg aatgccgttc     4980

tgaagctaaa tgtaaaaaga cgatatgata caacctatgg gacttacact ataaacatca     5040

cagttaattt aactgataag ggaaatattc agatatggtt accacagttc aaaagtaacg     5100

ctcgtgtcga tcttaacttg cgtccaactg gtggtggtac atatatcgga agaaattctg     5160

ttgatatgtg cttttatgat ggatatagta ctaacagcag ctctttggag ataagatttc     5220

aggatgataa ttctaaatct gatggaaaat tttatctaaa gaaaataaat gatgactcca     5280

aagaacttgt atacactttg tcacttctcc tggcaggtaa aaatttaaca ccaacaaatg     5340

gacaggcatt aaatattaac actgcttctc tggaaacaaa ctggaataga attacagctg     5400

tcaccatgcc agaaatcagt gttccggtgt tgtgttggcc tggacgtttg caattggatg     5460

caaaagtgaa aaatcccgag gctggacaat atatggggaa tattaaaatt actttcacac     5520

caagtagtca aacactctag tctagactcg agaccaggta ccggt                     5565


<210>  2
<211>  528
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Engineered E. coli cfaB gene with Lactococcus signal sequence

<400>  2
atgaaaaaaa agattatctc agctatttta atgtctacag tgatactttc tgctgcagcc       60

ccgttgtcag gtgtttacgc tgctagcgag aaaaatatta ctgtaacagc tagtgttgat      120

cctgcaattg atcttttgca agctgatggc aatgctctgc catcagctgt aaagttagct      180

tattctcccg catcaaaaac ttttgaaagt tacagagtaa tgactcaagt acatacaaac      240

gatgcaacta aaaaagtaat tgttaaactt gctgatacac cacagcttac agatgttctg      300

aattcaactg ttcaaatgcc tatcagtgtg tcatggggag gacaagtatt atctacaaca      360

gccaaagaat ttgaagctgc tgctttggga tattctgcat ccggtgtaaa tggcgtatca      420

tcttctcaag agttagtaat tagcgctgca cctaaaactg ccggtaccgc cccaactgca      480

ggaaactatt caggagtagt atctcttgta atgactttgg gatcctga                   528


<210>  3
<211>  642
<212>  DNA
<213>  artificial

<220>
<223>  Engineered E. coli cfaA gene with Lactococcus signal sequence

<400>  3
atgaaaaaga ttaatctggc attattgacc ctcgcaactt tgatgggtgt ttcttcaaca       60

gctgttgtat ttgccggcaa ctttatgata tatccaatat caaaagattt aaagaatgga      120

aatagcgagt tagttcgtgt ttattcaaaa tcaaaagaga tacaatatat aaaaatatat      180

acaaagaaga ttattaatcc cggtacaact gaagaatata aggttgatat acccaattgg      240

gatggtgggc ttgtagtcac tcctcagaaa gttatcctcc ctgcaggagc gagtaaatca      300

atacgtttaa ctcaatttaa aataccaaaa aaagaggaag tttacagagt atattttgag      360

gcggtaaaac cagatagcaa agaaaatgta attgataata aaaaactaac aacagagcta      420

tctgttaata taatttatgc ggctctaatt agatctttac caagtgaaca aaatatatca      480

ctaaatattt ctagaaatgc aaaaaaaaat ataattattt ataataatgg gaatgttaga      540

gcaggcgtta aagatattta tttttgtaag tcatctaata tcgatgataa ctgtgtaaaa      600

aaagcgtata acaagaatat atatccagaa aaagtcattt ga                         642


<210>  4
<211>  2607
<212>  DNA
<213>  artificial

<220>
<223>  Engineered E. coli cfaC gene with Lactococcus signal sequence

<400>  4
atgaaaaaga ttttgatcac tacgacatta gcacttgctc tcctgtcttt aggtgcagct       60

agcgttaccg gagatatacc cgactctttc cgtgatttat ggggagaaca agatgaattt      120

tatgaagtaa aactatatgg gcaaactcta ggaatacatc gaattaaaac aaccccaaca      180

catattaagt tttattcacc cgaaagcatt ttagataaaa taaatttaaa aaaagaaaag      240

gaaaaggaat tgagtgtttt ttttactaat tctttttcaa gaaatggcaa tatgagttgt      300

cagggtaaca ctactataca gtataactgc aattacatta aaacaaaatc agtagatgtc      360

atcgttgatg atgttgataa tgttgttaac ctttttatag gtaatgaatt tctggattct      420

gaagcacaca atgatgaata tcatcaatta tcacggaatg taaaaaaagc ttttatacaa      480

agccagacaa ttaatctctc agattctgga aagtataaaa gattgtctat ttcagggaat      540

agcgcgctgg gtattacaga tacaagttat gctgtcttaa attggtggat gaattacaat      600

aaatctaatg gttacagcaa caacgaaaaa acaatcaata gtttatactt tagacatgat      660

ttagataaga gatattatta tcaatttgga cgaatggatc gtacagattt atcacaaagt      720

attagcggga gctttaattt taacttactt cctttacccg atattgatgg tatacggaca      780

ggaaccacac aatcttatat caaaaataca gataagttta tcgcatcccc tgtaactgtt      840

atgttaacta atttttccag agtggaagct tttcgcaatg atcaattatt gggcgtatgg      900

tatttagatt ctggagtaaa tgaattagat acagctcgtt taccttatgg cagttacgat      960

cttaaattaa aaatttttga aaacactcaa ttagttcgtg aagaaataat tccttttaat     1020

aaaggaagaa gctctattgg tgatatgcaa tgggatattt tcgttcaggg agggaatatt     1080

gttaatgata atgatcgtta catagaaaaa caaaataatc ataagtcatc gattaatact     1140

gggctacgtt taccaattac gaaaaatatc tctgttcaac agggagtatc tgttatagat     1200

aataaaagtt attatgaagg aagtctgaaa tggaattccg gcattctatc tggctcacta     1260

aatagtgagt tcagttttct ttggggagat aatgcaaaag gtaattatca aagtatctcg     1320

tataccgatg gatttagctt atcattttat cataatgata agcgggtcga taattgtgga     1380

agaaattaca atgctggttg gagtggatgc tacgaatcat attcggcatc tttaagtatt     1440

cctttattag gatggacaag tactctggga tatagtgaca cttatagtga atcagtgtat     1500

aaaagccata ttctttctga atatggcttt tataatcaaa acatatataa agggagaacc     1560

caaagatggc aactgacttc atccacctct ttaaaatgga tggattataa ttttatgcca     1620

gcaattggaa tatataacag tgaacaaaga caactgactg ataaaggcgg atatatatct     1680

gtaactatca cccgagccag cagagaaaat tcattaaata cagggtattc ttacaactat     1740

tccagaggaa actattcttc taacgaatta tttgttgatg gatatatgac atcaacaaat     1800

aatggtgatt atcatgaggc aggaatgcgt tttaataaaa atagacataa tgcagaaggt     1860

agactttcag gtcgtataaa caatcgattt ggagatttaa atggttcatt cagcatgaat     1920

aaaaacagaa acaccaacag taccaatcat tctctcactg gtggttataa ttcctcattt     1980

gctcttacaa gtgatggatt ttactgggga ggaagtacag ctggtttgac aaaactggct     2040

ggcggtatta tcaaggttaa atcaaacgat actaaaaaaa acttggtaaa agtgactggg     2100

acattgtacg gtgattattc gctagggagc aacgataatg cttttattcc tgtaccagca     2160

ttaactccag ccagtttaat cattgaagat aataattatg gtgataataa tatttctata     2220

cttgcgccaa caaacaacga tatgtttatg ttgccgggta atgtttatcc tgttgaaatt     2280

gaaaccaaag taagtgtttc ttatattggt agaggttttg acccaaacgg cacgccactt     2340

tctggcgcac atgttttgaa tgaaccacat gttatcctgg atgaggacgg tggattttcg     2400

tttgaatata caggtaatga gaaaacactt tttttattaa agggcaggac tatttataca     2460

tgtcaactgg ggaaaaataa agttcataaa ggcattgttt tcgtcgggga tgttatatgt     2520

gatattaata gcacaagttc cttaccagat gaatttgtaa agaacccacg tgtgcaggat     2580

ttgctggcaa agaatgataa aggataa                                         2607


<210>  5
<211>  1119
<212>  DNA
<213>  artificial

<220>
<223>  Engineered E. coli cfaE gene with Lactococcus signal sequence

<400>  5
atgcaaagga aaaagaaagg gctatcgttc ttgttagccg gtacagtcgc tttaggggcg       60

ctggctgtct tgccagtcgg cgaaatccaa gcaaaggcgg ccgcagataa aaatcccgga      120

agtgaaaaca tgactaatac tattggtccc catgacaggg ggggatcttc ccccatatat      180

aatatcttaa attcctatct tacagcatac aatggaagcc atcatctgta tgataggatg      240

agttttttat gtttgtcttc tcaaaataca ctgaatggag catgcccaag cagtgatgcc      300

cctggcactg ctacaattga tggcgaaaca aatataacat tacaatttac ggaaaaaaga      360

agtctaatta aaagagaact gcaaattaaa ggctataaac aatttttgtt caaaaatgct      420

aattgcccat ctaaactagc acttaactca tctcattttc aatgtaatag agaacaagct      480

tcaggtgcta ctttatcgtt atacatacca gctggtgaat taaataaatt accttttggg      540

ggggtctgga atgccgttct gaagctaaat gtaaaaagac gatatgatac aacctatggg      600

acttacacta taaacatcac agttaattta actgataagg gaaatattca gatatggtta      660

ccacagttca aaagtaacgc tcgtgtcgat cttaacttgc gtccaactgg tggtggtaca      720

tatatcggaa gaaattctgt tgatatgtgc ttttatgatg gatatagtac taacagcagc      780

tctttggaga taagatttca ggatgataat tctaaatctg atggaaaatt ttatctaaag      840

aaaataaatg atgactccaa agaacttgta tacactttgt cacttctcct ggcaggtaaa      900

aatttaacac caacaaatgg acaggcatta aatattaaca ctgcttctct ggaaacaaac      960

tggaatagaa ttacagctgt caccatgcca gaaatcagtg ttccggtgtt gtgttggcct     1020

ggacgtttgc aattggatgc aaaagtgaaa aatcccgagg ctggacaata tatggggaat     1080

attaaaatta ctttcacacc aagtagtcaa acactctag                            1119


<210>  6
<211>  187
<212>  DNA
<213>  Lactococcus lactis

<400>  6
agtcttataa ctatactgac aatagaaaca ttaacaaatc taaaacagtc ttaattctat       60

cttgagaaag tattggtaat aatattattg tcgataacgc gagcataata aacggctctg      120

attaaattct gaagtttgtt agatacaatg atttcgttcg aaggaactac aaaataaatt      180

attctag                                                                187


<210>  7
<211>  142
<212>  DNA
<213>  Lactococcus lactis

<400>  7
gatatcaata tgcgaaaaga actatgaata tccactccat ttttggttgc catttgttaa       60

cgctgcctcc tctccctagt gctataataa aacaggccca ttttggaaca gacttctact      120

attttgttgt agatctgggc cc                                               142


<210>  8
<211>  194
<212>  DNA
<213>  Lactococcus lactis

<400>  8
catttggcag tttattcttg acatgtagtg agggggctgg tataatcaca tagtactgtt       60

tgattcttca gcaagactgg tacctcatga gagttataga ctcatggatc ttgctttgaa      120

gggttttgta cattataggc tcctatcaca tgctgaacct atggcctatt acattttttt      180

atatttcaag gagg                                                        194


<210>  9
<211>  175
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Engineered E. coli CfaB peptide with Lactococcus signal peptide

<400>  9

Met Lys Lys Lys Ile Ile Ser Ala Ile Leu Met Ser Thr Val Ile Leu 
1               5                   10                  15      


Ser Ala Ala Ala Pro Leu Ser Gly Val Tyr Ala Ala Ser Glu Lys Asn 
            20                  25                  30          


Ile Thr Val Thr Ala Ser Val Asp Pro Ala Ile Asp Leu Leu Gln Ala 
        35                  40                  45              


Asp Gly Asn Ala Leu Pro Ser Ala Val Lys Leu Ala Tyr Ser Pro Ala 
    50                  55                  60                  


Ser Lys Thr Phe Glu Ser Tyr Arg Val Met Thr Gln Val His Thr Asn 
65                  70                  75                  80  


Asp Ala Thr Lys Lys Val Ile Val Lys Leu Ala Asp Thr Pro Gln Leu 
                85                  90                  95      


Thr Asp Val Leu Asn Ser Thr Val Gln Met Pro Ile Ser Val Ser Trp 
            100                 105                 110         


Gly Gly Gln Val Leu Ser Thr Thr Ala Lys Glu Phe Glu Ala Ala Ala 
        115                 120                 125             


Leu Gly Tyr Ser Ala Ser Gly Val Asn Gly Val Ser Ser Ser Gln Glu 
    130                 135                 140                 


Leu Val Ile Ser Ala Ala Pro Lys Thr Ala Gly Thr Ala Pro Thr Ala 
145                 150                 155                 160 


Gly Asn Tyr Ser Gly Val Val Ser Leu Val Met Thr Leu Gly Ser 
                165                 170                 175 


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

<220>
<223>  Engineered E. coli CfaA peptide with Lactococcus signal peptide

<400>  10

Met Lys Lys Ile Asn Leu Ala Leu Leu Thr Leu Ala Thr Leu Met Gly 
1               5                   10                  15      


Val Ser Ser Thr Ala Val Val Phe Ala Gly Asn Phe Met Ile Tyr Pro 
            20                  25                  30          


Ile Ser Lys Asp Leu Lys Asn Gly Asn Ser Glu Leu Val Arg Val Tyr 
        35                  40                  45              


Ser Lys Ser Lys Glu Ile Gln Tyr Ile Lys Ile Tyr Thr Lys Lys Ile 
    50                  55                  60                  


Ile Asn Pro Gly Thr Thr Glu Glu Tyr Lys Val Asp Ile Pro Asn Trp 
65                  70                  75                  80  


Asp Gly Gly Leu Val Val Thr Pro Gln Lys Val Ile Leu Pro Ala Gly 
                85                  90                  95      


Ala Ser Lys Ser Ile Arg Leu Thr Gln Phe Lys Ile Pro Lys Lys Glu 
            100                 105                 110         


Glu Val Tyr Arg Val Tyr Phe Glu Ala Val Lys Pro Asp Ser Lys Glu 
        115                 120                 125             


Asn Val Ile Asp Asn Lys Lys Leu Thr Thr Glu Leu Ser Val Asn Ile 
    130                 135                 140                 


Ile Tyr Ala Ala Leu Ile Arg Ser Leu Pro Ser Glu Gln Asn Ile Ser 
145                 150                 155                 160 


Leu Asn Ile Ser Arg Asn Ala Lys Lys Asn Ile Ile Ile Tyr Asn Asn 
                165                 170                 175     


Gly Asn Val Arg Ala Gly Val Lys Asp Ile Tyr Phe Cys Lys Ser Ser 
            180                 185                 190         


Asn Ile Asp Asp Asn Cys Val Lys Lys Ala Tyr Asn Lys Asn Ile Tyr 
        195                 200                 205             


Pro Glu Lys Val Ile 
    210             


<210>  11
<211>  868
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Engineered E. coli CfaC peptide with Lactococcus signal peptide

<400>  11

Met Lys Lys Ile Leu Ile Thr Thr Thr Leu Ala Leu Ala Leu Leu Ser 
1               5                   10                  15      


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


Leu Trp Gly Glu Gln Asp Glu Phe Tyr Glu Val Lys Leu Tyr Gly Gln 
        35                  40                  45              


Thr Leu Gly Ile His Arg Ile Lys Thr Thr Pro Thr His Ile Lys Phe 
    50                  55                  60                  


Tyr Ser Pro Glu Ser Ile Leu Asp Lys Ile Asn Leu Lys Lys Glu Lys 
65                  70                  75                  80  


Glu Lys Glu Leu Ser Val Phe Phe Thr Asn Ser Phe Ser Arg Asn Gly 
                85                  90                  95      


Asn Met Ser Cys Gln Gly Asn Thr Thr Ile Gln Tyr Asn Cys Asn Tyr 
            100                 105                 110         


Ile Lys Thr Lys Ser Val Asp Val Ile Val Asp Asp Val Asp Asn Val 
        115                 120                 125             


Val Asn Leu Phe Ile Gly Asn Glu Phe Leu Asp Ser Glu Ala His Asn 
    130                 135                 140                 


Asp Glu Tyr His Gln Leu Ser Arg Asn Val Lys Lys Ala Phe Ile Gln 
145                 150                 155                 160 


Ser Gln Thr Ile Asn Leu Ser Asp Ser Gly Lys Tyr Lys Arg Leu Ser 
                165                 170                 175     


Ile Ser Gly Asn Ser Ala Leu Gly Ile Thr Asp Thr Ser Tyr Ala Val 
            180                 185                 190         


Leu Asn Trp Trp Met Asn Tyr Asn Lys Ser Asn Gly Tyr Ser Asn Asn 
        195                 200                 205             


Glu Lys Thr Ile Asn Ser Leu Tyr Phe Arg His Asp Leu Asp Lys Arg 
    210                 215                 220                 


Tyr Tyr Tyr Gln Phe Gly Arg Met Asp Arg Thr Asp Leu Ser Gln Ser 
225                 230                 235                 240 


Ile Ser Gly Ser Phe Asn Phe Asn Leu Leu Pro Leu Pro Asp Ile Asp 
                245                 250                 255     


Gly Ile Arg Thr Gly Thr Thr Gln Ser Tyr Ile Lys Asn Thr Asp Lys 
            260                 265                 270         


Phe Ile Ala Ser Pro Val Thr Val Met Leu Thr Asn Phe Ser Arg Val 
        275                 280                 285             


Glu Ala Phe Arg Asn Asp Gln Leu Leu Gly Val Trp Tyr Leu Asp Ser 
    290                 295                 300                 


Gly Val Asn Glu Leu Asp Thr Ala Arg Leu Pro Tyr Gly Ser Tyr Asp 
305                 310                 315                 320 


Leu Lys Leu Lys Ile Phe Glu Asn Thr Gln Leu Val Arg Glu Glu Ile 
                325                 330                 335     


Ile Pro Phe Asn Lys Gly Arg Ser Ser Ile Gly Asp Met Gln Trp Asp 
            340                 345                 350         


Ile Phe Val Gln Gly Gly Asn Ile Val Asn Asp Asn Asp Arg Tyr Ile 
        355                 360                 365             


Glu Lys Gln Asn Asn His Lys Ser Ser Ile Asn Thr Gly Leu Arg Leu 
    370                 375                 380                 


Pro Ile Thr Lys Asn Ile Ser Val Gln Gln Gly Val Ser Val Ile Asp 
385                 390                 395                 400 


Asn Lys Ser Tyr Tyr Glu Gly Ser Leu Lys Trp Asn Ser Gly Ile Leu 
                405                 410                 415     


Ser Gly Ser Leu Asn Ser Glu Phe Ser Phe Leu Trp Gly Asp Asn Ala 
            420                 425                 430         


Lys Gly Asn Tyr Gln Ser Ile Ser Tyr Thr Asp Gly Phe Ser Leu Ser 
        435                 440                 445             


Phe Tyr His Asn Asp Lys Arg Val Asp Asn Cys Gly Arg Asn Tyr Asn 
    450                 455                 460                 


Ala Gly Trp Ser Gly Cys Tyr Glu Ser Tyr Ser Ala Ser Leu Ser Ile 
465                 470                 475                 480 


Pro Leu Leu Gly Trp Thr Ser Thr Leu Gly Tyr Ser Asp Thr Tyr Ser 
                485                 490                 495     


Glu Ser Val Tyr Lys Ser His Ile Leu Ser Glu Tyr Gly Phe Tyr Asn 
            500                 505                 510         


Gln Asn Ile Tyr Lys Gly Arg Thr Gln Arg Trp Gln Leu Thr Ser Ser 
        515                 520                 525             


Thr Ser Leu Lys Trp Met Asp Tyr Asn Phe Met Pro Ala Ile Gly Ile 
    530                 535                 540                 


Tyr Asn Ser Glu Gln Arg Gln Leu Thr Asp Lys Gly Gly Tyr Ile Ser 
545                 550                 555                 560 


Val Thr Ile Thr Arg Ala Ser Arg Glu Asn Ser Leu Asn Thr Gly Tyr 
                565                 570                 575     


Ser Tyr Asn Tyr Ser Arg Gly Asn Tyr Ser Ser Asn Glu Leu Phe Val 
            580                 585                 590         


Asp Gly Tyr Met Thr Ser Thr Asn Asn Gly Asp Tyr His Glu Ala Gly 
        595                 600                 605             


Met Arg Phe Asn Lys Asn Arg His Asn Ala Glu Gly Arg Leu Ser Gly 
    610                 615                 620                 


Arg Ile Asn Asn Arg Phe Gly Asp Leu Asn Gly Ser Phe Ser Met Asn 
625                 630                 635                 640 


Lys Asn Arg Asn Thr Asn Ser Thr Asn His Ser Leu Thr Gly Gly Tyr 
                645                 650                 655     


Asn Ser Ser Phe Ala Leu Thr Ser Asp Gly Phe Tyr Trp Gly Gly Ser 
            660                 665                 670         


Thr Ala Gly Leu Thr Lys Leu Ala Gly Gly Ile Ile Lys Val Lys Ser 
        675                 680                 685             


Asn Asp Thr Lys Lys Asn Leu Val Lys Val Thr Gly Thr Leu Tyr Gly 
    690                 695                 700                 


Asp Tyr Ser Leu Gly Ser Asn Asp Asn Ala Phe Ile Pro Val Pro Ala 
705                 710                 715                 720 


Leu Thr Pro Ala Ser Leu Ile Ile Glu Asp Asn Asn Tyr Gly Asp Asn 
                725                 730                 735     


Asn Ile Ser Ile Leu Ala Pro Thr Asn Asn Asp Met Phe Met Leu Pro 
            740                 745                 750         


Gly Asn Val Tyr Pro Val Glu Ile Glu Thr Lys Val Ser Val Ser Tyr 
        755                 760                 765             


Ile Gly Arg Gly Phe Asp Pro Asn Gly Thr Pro Leu Ser Gly Ala His 
    770                 775                 780                 


Val Leu Asn Glu Pro His Val Ile Leu Asp Glu Asp Gly Gly Phe Ser 
785                 790                 795                 800 


Phe Glu Tyr Thr Gly Asn Glu Lys Thr Leu Phe Leu Leu Lys Gly Arg 
                805                 810                 815     


Thr Ile Tyr Thr Cys Gln Leu Gly Lys Asn Lys Val His Lys Gly Ile 
            820                 825                 830         


Val Phe Val Gly Asp Val Ile Cys Asp Ile Asn Ser Thr Ser Ser Leu 
        835                 840                 845             


Pro Asp Glu Phe Val Lys Asn Pro Arg Val Gln Asp Leu Leu Ala Lys 
    850                 855                 860                 


Asn Asp Lys Gly 
865             


<210>  12
<211>  372
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Engineered E. coli CfaE peptide with Lactococcus signal peptide

<400>  12

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


Ala Leu Gly Ala Leu Ala Val Leu Pro Val Gly Glu Ile Gln Ala Lys 
            20                  25                  30          


Ala Ala Ala Asp Lys Asn Pro Gly Ser Glu Asn Met Thr Asn Thr Ile 
        35                  40                  45              


Gly Pro His Asp Arg Gly Gly Ser Ser Pro Ile Tyr Asn Ile Leu Asn 
    50                  55                  60                  


Ser Tyr Leu Thr Ala Tyr Asn Gly Ser His His Leu Tyr Asp Arg Met 
65                  70                  75                  80  


Ser Phe Leu Cys Leu Ser Ser Gln Asn Thr Leu Asn Gly Ala Cys Pro 
                85                  90                  95      


Ser Ser Asp Ala Pro Gly Thr Ala Thr Ile Asp Gly Glu Thr Asn Ile 
            100                 105                 110         


Thr Leu Gln Phe Thr Glu Lys Arg Ser Leu Ile Lys Arg Glu Leu Gln 
        115                 120                 125             


Ile Lys Gly Tyr Lys Gln Phe Leu Phe Lys Asn Ala Asn Cys Pro Ser 
    130                 135                 140                 


Lys Leu Ala Leu Asn Ser Ser His Phe Gln Cys Asn Arg Glu Gln Ala 
145                 150                 155                 160 


Ser Gly Ala Thr Leu Ser Leu Tyr Ile Pro Ala Gly Glu Leu Asn Lys 
                165                 170                 175     


Leu Pro Phe Gly Gly Val Trp Asn Ala Val Leu Lys Leu Asn Val Lys 
            180                 185                 190         


Arg Arg Tyr Asp Thr Thr Tyr Gly Thr Tyr Thr Ile Asn Ile Thr Val 
        195                 200                 205             


Asn Leu Thr Asp Lys Gly Asn Ile Gln Ile Trp Leu Pro Gln Phe Lys 
    210                 215                 220                 


Ser Asn Ala Arg Val Asp Leu Asn Leu Arg Pro Thr Gly Gly Gly Thr 
225                 230                 235                 240 


Tyr Ile Gly Arg Asn Ser Val Asp Met Cys Phe Tyr Asp Gly Tyr Ser 
                245                 250                 255     


Thr Asn Ser Ser Ser Leu Glu Ile Arg Phe Gln Asp Asp Asn Ser Lys 
            260                 265                 270         


Ser Asp Gly Lys Phe Tyr Leu Lys Lys Ile Asn Asp Asp Ser Lys Glu 
        275                 280                 285             


Leu Val Tyr Thr Leu Ser Leu Leu Leu Ala Gly Lys Asn Leu Thr Pro 
    290                 295                 300                 


Thr Asn Gly Gln Ala Leu Asn Ile Asn Thr Ala Ser Leu Glu Thr Asn 
305                 310                 315                 320 


Trp Asn Arg Ile Thr Ala Val Thr Met Pro Glu Ile Ser Val Pro Val 
                325                 330                 335     


Leu Cys Trp Pro Gly Arg Leu Gln Leu Asp Ala Lys Val Lys Asn Pro 
            340                 345                 350         


Glu Ala Gly Gln Tyr Met Gly Asn Ile Lys Ile Thr Phe Thr Pro Ser 
        355                 360                 365             


Ser Gln Thr Leu 
    370         


