3
Rust et al., Figure S1 A B Amino acid sequence comparison and alignment of various bacterial FlgM proteins color-coded according to different physico-chemical properties. Sequence alignments were performed using ClustalW2 (Larkin et al., 2007); the alignment was color-coded and depicted using the software GeneDoc (Nicholas et al., 1997). A) alignment of FlgM of different Campylobacterales species which all possess short N-termini; B) alignment of FlgM of diverse eubacteria. H . p y l o r i _ 2 6 6 9 5 : H . p y l o r i _ J 9 9 : H . h e p a t i c u s : H P A G 1 _ 1 0 6 0 : H . a c i n o n y c h i s : H . m u s t e l a e : C . j e j u n i : W . s u c c i n o g e n e s : B . s u b t i l i s : A . a e o l i c u s : E . c o l i : S . T y p h i m u r i u m : Y . e n t e r o c o l i t i c a : P . m i r a b i l i s : V . c h o l e r a e : P . p u t i d a : B . b a c t e r i o v o r u s : * 2 0 * 4 0 * 6 0 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M N I K L K D F T M I N A V S S L A P V Q S L G N Y K R V E K - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M I N A V S S L T P V Q S V G N Y K R V E K - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M V N G V N T S V K V A N M L N R D V L A K - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M N I K L K D F T M I N A V S S L A P V Q S L G N Y K R V E K - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M I N A I S S L T P T Q S W G N Y K R V E K - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M L N I I G K N P M V A N V D S K I H K V G - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M I N P I Q Q S Y V A N T A L N T N R I D K - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M I S P I Q S G A T L P T A K N V Q N R E V - - - - - - - - - - - - - - - - - M K I N Q F G T Q S V N P Y Q K N Y D K Q A V Q K T V A Q P Q D K I E I S S Q A K E M - - - - - - - - - - - - - - - - - - M V N R I E L S R L I G L L L E T E K R K N T E Q K E S G T N K I E D K V T L S K I - - - - - - - - - - - - M S I D R T S P L K P V S T V Q P R E T T D A P V T N T R - - - A A K T T A S T S T S V T L S D - - - - - - - - - - - - M S I D R T S P L K P V S T V Q T R E T S D T P V Q K T R - - - Q E K T S A A T S A S V T L S D - - - - - - - - - - - - M S I D R T Q P L L P V T P V Q P R E T S D I A Q Q T R K P - S A Q S K T P V S G T E V K L S D - - - - - - - - - - - - M S I E R T N P L I P I T A I S Q R N L N E G A Q E A R K T G N A Q T K A M A G D T S V K L S E - - - - M A G I D N I R A G Q S L N T T S R T N V R S S D A S N S S T Q S G A V S R S S A S G Q D A V S L S S Q S K A I - - - - M V I D F S R L N N S P S V T G G V R G N T A S G S A E K P A A S Q E A V K D T S A S G E A V H L S Q E A Q Q L K I T H N K V G Q N L N L T D S S R A D N A A G I K G K A D N I G A A K A D V L T S S N L G E S S R V E L S P R A Q E A : 3 1 : 2 2 : 2 2 : 3 1 : 2 2 : 2 2 : 2 2 : 2 2 : 4 3 : 4 2 : 4 5 : 4 5 : 4 7 : 4 8 : 5 6 : 5 6 : 6 0 H . p y l o r i _ 2 6 6 9 5 : H . p y l o r i _ J 9 9 : H . h e p a t i c u s : H P A G 1 _ 1 0 6 0 : H . a c i n o n y c h i s : H . m u s t e l a e : C . j e j u n i : W . s u c c i n o g e n e s : B . s u b t i l i s : A . a e o l i c u s : E . c o l i : S . T y p h i m u r i u m : Y . e n t e r o c o l i t i c a : P . m i r a b i l i s : V . c h o l e r a e : P . p u t i d a : B . b a c t e r i o v o r u s : * 8 0 * 1 0 0 * N E K V E N N E - - - - A A L D R V A E I K K A I E N N Q Y K I N L H E T S H K M A K D L L G I S - - - N E K V E N N E - - - - A A L D R V A E I K Q A I E N N Q Y K I N L H E T S H K M A Q D L L G I S - - - Q N E N N E V K E K E Q T Q L S K V E Q I K E Q I K N G E Y K I D L Q Q T S E K M A S N L L N L - - - - N E K V E N S E - - - - A A L D R V A E I K K A I E N N Q Y K I N L H E T S H K M A Q D L L G I S - - - N E E V Q N N E - - - - V A L D K V A Q I K H A I E N N Q Y K I N L R E T S H K M A Q D L L G I S - - - D Q K V Q A E K - - - - I Q S D K V E K I S Q A V K S G G Y K V D L Q A T S E K M A L N L L G L - - - - E T K T N D T Q - - - K A E N D K A S K I A E Q I K N G T Y K I D T K A T A A A I A D S L I - - - - - - E S K T E S S - - - - - A S S S K V Q A I K E S L Q K G E Y K L D L P A T A E K M A L N L L N R E - - - Q H A S D A V T G S - - - R Q E K I A Q L K A Q I E N G S Y K V D A N H I A K N M I N F Y K K Q - - - - A Q E L S K N D V E E K D L E K K V K E L K E K I E K G E Y E V S D E K V V K G L I E F F T - - - - - - A Q A K L M Q P G S S D I N L E R V E V L K L A I R N G E L K M D T G K I A D A L I N E A Q Q D L Q S N A Q A K L M Q P G V S D I N M E R V E A L K T A I R N G E L K M D T G K I A D S L I R E A Q S Y L Q S K A Q A K L M Q P G S Q D I N V E R V E T L K Q A I R S G Q L T M D T G K I A D A L L K N V A D D L K N S A Q K K L V Q P S N Q D I N V E K V A R L K A A I A D G T L T M D S S K I A E A L F R E A A E S I T K - G E M H S E M A S R P S F D S A K V A A I K E A I A N G S Y V V D P E K L A D N M I K F E K E L G G F - Q K I S D K L R D E P V V N S A R V A Q L K Q A I A D G S Y Q V D A G R V A S K L L D F E A Q R - - - - K R I K E L A L A T P D V D E A K V A K F R A L I D E G K Y K V D A K A I A D K M V D E H L E F - - - - 4 6 6 6 : 7 6 : 6 7 : 7 0 : 7 6 : 6 7 : 6 6 : 6 5 : 6 6 : 8 8 : 8 8 : 9 7 : 9 7 : 9 9 : 9 9 : 1 0 7 : 1 0 4 : 1 0 8 H . p y l o r i 2 6 6 9 5 : H P A G 1 _ 1 0 6 0 : H . p y l o r i _ J 9 9 : H . a c i n o n y c h i s : H . m u s t e l a e : H . h e p a t i c u s : W . s u c c i n o g e n e s : C . j e j u n i : * 2 0 * 4 0 * 6 0 * 8 0 M N I K L K D F T M I N A V S S L A P V Q S L G N Y K R V E K N E K V E N N E - - - - A A L D R V A E I K K A I E N N Q Y K I N L H E T S H K M A K D L L G I S M N I K L K D F T M I N A V S S L A P V Q S L G N Y K R V E K N E K V E N S E - - - - A A L D R V A E I K K A I E N N Q Y K I N L H E T S H K M A Q D L L G I S - - - - - - - - - M I N A V S S L T P V Q S V G N Y K R V E K N E K V E N N E - - - - A A L D R V A E I K Q A I E N N Q Y K I N L H E T S H K M A Q D L L G I S - - - - - - - - - M I N A I S S L T P T Q S W G N Y K R V E K N E E V Q N N E - - - - V A L D K V A Q I K H A I E N N Q Y K I N L R E T S H K M A Q D L L G I S - - - - - - - - - M L N I I G K N P M V A N V D S K I H K V G D Q K V Q A E K - - - - I Q S D K V E K I S Q A V K S G G Y K V D L Q A T S E K M A L N L L G L - - - - - - - - - - M V N G V N T S V K V A N M L N R D V L A K Q N E N N E V K E K E Q T Q L S K V E Q I K E Q I K N G E Y K I D L Q Q T S E K M A S N L L N L - - - - - - - - - - M I S P I Q S G A T L P T A K N V Q N R E V E S K T E S S - - - - - A S S S K V Q A I K E S L Q K G E Y K L D L P A T A E K M A L N L L N R E - - - - - - - - - M I N P I Q Q S Y V A N T A L N T N R I D K E T K T N D T Q - - - K A E N D K A S K I A E Q I K N G T Y K I D T K A T A A A I A D S L I - - - M 6 n 6 n k k d 4 v I k 6 n Y K 6 1 l T s k 6 A L 6 : 7 6 : 7 6 : 6 7 : 6 7 : 6 6 : 7 0 : 6 6 : 6 5

Rust et al., Figure S1

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Page 1: Rust et al., Figure S1

Rust et al., Figure S1

A

B

Amino acid sequence comparison and alignment of various bacterial FlgM proteins color-coded according to different physico-chemical properties. Sequence alignments were performed using ClustalW2 (Larkin et al., 2007); the alignment was color-coded and depicted using the software GeneDoc (Nicholas et al., 1997). A) alignment of FlgM of different Campylobacterales species which all possess short N-termini; B) alignment of FlgM of diverse eubacteria.

H.pylori_26695 : H.pylori_J99 : H.hepaticus : HPAG1_1060 : H.acinonychis : H.mustelae : C.jejuni : W.succinogenes : B.subtilis : A.aeolicus : E.coli : S.Typhimurium : Y.enterocolitica : P.mirabilis : V.cholerae : P.putida : B.bacteriovorus :

* 20 * 40 * 60-----------------------------MNIKLKDFTMINAVSSLAPVQSLGNYKRVEK--------------------------------------MINAVSSLTPVQSVGNYKRVEK--------------------------------------MVNGVNTSVKVANMLNRDVLAK-----------------------------MNIKLKDFTMINAVSSLAPVQSLGNYKRVEK--------------------------------------MINAISSLTPTQSWGNYKRVEK--------------------------------------MLNIIGKNPMVANVDSKIHKVG--------------------------------------MINPIQQSYVANTALNTNRIDK--------------------------------------MISPIQSGATLPTAKNVQNREV-----------------MKINQFGTQSVNPYQKNYDKQAVQKTVAQPQDKIEISSQAKEM------------------MVNRIELSRLIGLLLETEKRKNTEQKESGTNKIEDKVTLSKI------------MSIDRTSPLKPVSTVQPRETTDAPVTNTR---AAKTTASTSTSVTLSD------------MSIDRTSPLKPVSTVQTRETSDTPVQKTR---QEKTSAATSASVTLSD------------MSIDRTQPLLPVTPVQPRETSDIAQQTRKP-SAQSKTPVSGTEVKLSD------------MSIERTNPLIPITAISQRNLNEGAQEARKTGNAQTKAMAGDTSVKLSE----MAGIDNIRAGQSLNTTSRTNVRSSDASNSSTQSGAVSRSSASGQDAVSLSSQSKAI----MVIDFSRLNNSPSVTGGVRGNTASGSAEKPAASQEAVKDTSASGEAVHLSQEAQQLKITHNKVGQNLNLTDSSRADNAAGIKGKADNIGAAKADVLTSSNLGESSRVELSPRAQEA

: 31 : 22 : 22 : 31 : 22 : 22 : 22 : 22 : 43 : 42 : 45 : 45 : 47 : 48 : 56 : 56 : 60

H.pylori_26695 : H.pylori_J99 : H.hepaticus : HPAG1_1060 : H.acinonychis : H.mustelae : C.jejuni : W.succinogenes : B.subtilis : A.aeolicus : E.coli : S.Typhimurium : Y.enterocolitica : P.mirabilis : V.cholerae : P.putida : B.bacteriovorus :

* 80 * 100 * NEKVENNE----AALDRVAEIKKAIENNQYKINLHETSHKMAKDLLGIS---NEKVENNE----AALDRVAEIKQAIENNQYKINLHETSHKMAQDLLGIS---QNENNEVKEKEQTQLSKVEQIKEQIKNGEYKIDLQQTSEKMASNLLNL----NEKVENSE----AALDRVAEIKKAIENNQYKINLHETSHKMAQDLLGIS---NEEVQNNE----VALDKVAQIKHAIENNQYKINLRETSHKMAQDLLGIS---DQKVQAEK----IQSDKVEKISQAVKSGGYKVDLQATSEKMALNLLGL----ETKTNDTQ---KAENDKASKIAEQIKNGTYKIDTKATAAAIADSLI------ESKTESS-----ASSSKVQAIKESLQKGEYKLDLPATAEKMALNLLNRE---QHASDAVTGS---RQEKIAQLKAQIENGSYKVDANHIAKNMINFYKKQ----AQELSKNDVEEKDLEKKVKELKEKIEKGEYEVSDEKVVKGLIEFFT------AQAKLMQPGSSDINLERVEVLKLAIRNGELKMDTGKIADALINEAQQDLQSNAQAKLMQPGVSDINMERVEALKTAIRNGELKMDTGKIADSLIREAQSYLQSKAQAKLMQPGSQDINVERVETLKQAIRSGQLTMDTGKIADALLKNVADDLKNSAQKKLVQPSNQDINVEKVARLKAAIADGTLTMDSSKIAEALFREAAESITK-GEMHSEMASRPSFDSAKVAAIKEAIANGSYVVDPEKLADNMIKFEKELGGF-QKISDKLRDEPVVNSARVAQLKQAIADGSYQVDAGRVASKLLDFEAQR----KRIKELALATPDVDEAKVAKFRALIDEGKYKVDAKAIADKMVDEHLEF---- 4 6 6 6

: 76 : 67 : 70 : 76 : 67 : 66 : 65 : 66 : 88 : 88 : 97 : 97 : 99 : 99 : 107 : 104 : 108

H.pylori26695 : HPAG1_1060 : H.pylori_J99 : H.acinonychis : H.mustelae : H.hepaticus : W.succinogenes : C.jejuni :

* 20 * 40 * 60 * 80MNIKLKDFTMINAVSSLAPVQSLGNYKRVEKNEKVENNE----AALDRVAEIKKAIENNQYKINLHETSHKMAKDLLGISMNIKLKDFTMINAVSSLAPVQSLGNYKRVEKNEKVENSE----AALDRVAEIKKAIENNQYKINLHETSHKMAQDLLGIS---------MINAVSSLTPVQSVGNYKRVEKNEKVENNE----AALDRVAEIKQAIENNQYKINLHETSHKMAQDLLGIS---------MINAISSLTPTQSWGNYKRVEKNEEVQNNE----VALDKVAQIKHAIENNQYKINLRETSHKMAQDLLGIS---------MLNIIGKNPMVANVDSKIHKVGDQKVQAEK----IQSDKVEKISQAVKSGGYKVDLQATSEKMALNLLGL----------MVNGVNTSVKVANMLNRDVLAKQNENNEVKEKEQTQLSKVEQIKEQIKNGEYKIDLQQTSEKMASNLLNL----------MISPIQSGATLPTAKNVQNREVESKTESS-----ASSSKVQAIKESLQKGEYKLDLPATAEKMALNLLNRE---------MINPIQQSYVANTALNTNRIDKETKTNDTQ---KAENDKASKIAEQIKNGTYKIDTKATAAAIADSLI--- M6n 6 n k k d4v Ik 6 n YK61l Ts k6A L6

: 76 : 76 : 67 : 67 : 66 : 70 : 66 : 65

Page 2: Rust et al., Figure S1

Rust et al., Figure S2

Amino acid sequence alignment and comparison of H. pylori FliA and FlhA (C-terminal cytoplasmic domain only). Alignment was color-coded according to different physico-chemical properties. The sequence alignment was performed using ClustalW2 (Larkin et al., 2007); the alignment was color-coded and depicted using the software GeneDoc (Nicholas et al., 1997).

HPFlhA : HPFliA :

* 20 * 40 * 60 *ASDYGFLMPQIRIRDNLQLPPTHYEIKLKGIVIGEGMVMPDKFLAMNTGFVNKEIEGIPTKEPAFGMDAL---------------------------------------------MILMMENRMPKGIQKTETSEKNIEK M N4 GI E

: 70 : 25

HPFlhA : HPFliA :

80 * 100 * 120 * 140WIETKNKEEAIIQGYTIIDPSTVIATHTSELVKKYAEDFITKDEVKSLLERLAKDYPTIVEESKKIPTGAVLNAYDKQQHHHQDDLAIQYLPAVRAMAFRLKERLPSSIDFNDLVSIGTEELIKLARRYESALNDSFWGY 6 1K22 Q I 6 L 4 D V E L K G

: 140 : 95

HPFlhA : HPFliA :

* 160 * 180 * 200 *IRSVLQALLHEKIPIKDMLTILETITDIAPLVQNDVNILTEQVRARLSRVITNAFKSEDGRLKFLTFSTDAKTRVNGAMLDYLRSLDVISR-SSRKLIKSIDIEITKHLNEHGKEPSDAYLAQTLGENIEKIKEAKTASD 43 6 6 6 D663 3 I 6 L E 4 6 46K 3D

: 210 : 164

HPFlhA : HPFliA :

220 * 240 * 260 * 280SEQFLLNKLRENGTSKSLLLNVGELQKLIEVVSEEAMKVLQKGIAPVILIVEPNLRKALSNQMEQARIDVIYALVPIDEQFNAIEQDEITKKIEAEELLEHVQKALNQMSEREQILIQLYYFEELNLSEIKEILGITESR 6 N 6 E 2 L6E V 6 24 6 L L 26

: 280 : 234

HPFlhA : HPFliA :

* 300 IVLSHAELDPNSNFEALGTIHINFISQIIKEVIKKVR-KSLGVDHG--I E6 LG H

: 304 : 255

HPFlhA : HPFliA :

* 20 * 40 * 60 *ASDYGFLMPQIRIRDNLQLPPTHYEIKLKGIVIGEGMVMPDKFLAMNTGFVNKEIEGIPTKEPAFGMDAL---------------------------------------------MILMMENRMPKGIQKTETSEKNIEK M N4 GI E

: 70 : 25

HPFlhA : HPFliA :

80 * 100 * 120 * 140WIETKNKEEAIIQGYTIIDPSTVIATHTSELVKKYAEDFITKDEVKSLLERLAKDYPTIVEESKKIPTGAVLNAYDKQQHHHQDDLAIQYLPAVRAMAFRLKERLPSSIDFNDLVSIGTEELIKLARRYESALNDSFWGY 6 1K22 Q I 6 L 4 D V E L K G

: 140 : 95

HPFlhA : HPFliA :

* 160 * 180 * 200 *IRSVLQALLHEKIPIKDMLTILETITDIAPLVQNDVNILTEQVRARLSRVITNAFKSEDGRLKFLTFSTDAKTRVNGAMLDYLRSLDVISR-SSRKLIKSIDIEITKHLNEHGKEPSDAYLAQTLGENIEKIKEAKTASD 43 6 6 6 D663 3 I 6 L E 4 6 46K 3D

: 210 : 164

HPFlhA : HPFliA :

220 * 240 * 260 * 280SEQFLLNKLRENGTSKSLLLNVGELQKLIEVVSEEAMKVLQKGIAPVILIVEPNLRKALSNQMEQARIDVIYALVPIDEQFNAIEQDEITKKIEAEELLEHVQKALNQMSEREQILIQLYYFEELNLSEIKEILGITESR 6 N 6 E 2 L6E V 6 24 6 L L 26

: 280 : 234

HPFlhA : HPFliA :

* 300 IVLSHAELDPNSNFEALGTIHINFISQIIKEVIKKVR-KSLGVDHG--I E6 LG H

: 304 : 255

HPFlhA : HPFliA :

* 20 * 40 * 60 *ASDYGFLMPQIRIRDNLQLPPTHYEIKLKGIVIGEGMVMPDKFLAMNTGFVNKEIEGIPTKEPAFGMDAL---------------------------------------------MILMMENRMPKGIQKTETSEKNIEK M N4 GI E

: 70 : 25

HPFlhA : HPFliA :

80 * 100 * 120 * 140WIETKNKEEAIIQGYTIIDPSTVIATHTSELVKKYAEDFITKDEVKSLLERLAKDYPTIVEESKKIPTGAVLNAYDKQQHHHQDDLAIQYLPAVRAMAFRLKERLPSSIDFNDLVSIGTEELIKLARRYESALNDSFWGY 6 1K22 Q I 6 L 4 D V E L K G

: 140 : 95

HPFlhA : HPFliA :

* 160 * 180 * 200 *IRSVLQALLHEKIPIKDMLTILETITDIAPLVQNDVNILTEQVRARLSRVITNAFKSEDGRLKFLTFSTDAKTRVNGAMLDYLRSLDVISR-SSRKLIKSIDIEITKHLNEHGKEPSDAYLAQTLGENIEKIKEAKTASD 43 6 6 6 D663 3 I 6 L E 4 6 46K 3D

: 210 : 164

HPFlhA : HPFliA :

220 * 240 * 260 * 280SEQFLLNKLRENGTSKSLLLNVGELQKLIEVVSEEAMKVLQKGIAPVILIVEPNLRKALSNQMEQARIDVIYALVPIDEQFNAIEQDEITKKIEAEELLEHVQKALNQMSEREQILIQLYYFEELNLSEIKEILGITESR 6 N 6 E 2 L6E V 6 24 6 L L 26

: 280 : 234

HPFlhA : HPFliA :

* 300 IVLSHAELDPNSNFEALGTIHINFISQIIKEVIKKVR-KSLGVDHG--I E6 LG H

: 304 : 255

HPFlhA : HPFliA :

* 20 * 40 * 60 *ASDYGFLMPQIRIRDNLQLPPTHYEIKLKGIVIGEGMVMPDKFLAMNTGFVNKEIEGIPTKEPAFGMDAL---------------------------------------------MILMMENRMPKGIQKTETSEKNIEK M N4 GI E

: 70 : 25

HPFlhA : HPFliA :

80 * 100 * 120 * 140WIETKNKEEAIIQGYTIIDPSTVIATHTSELVKKYAEDFITKDEVKSLLERLAKDYPTIVEESKKIPTGAVLNAYDKQQHHHQDDLAIQYLPAVRAMAFRLKERLPSSIDFNDLVSIGTEELIKLARRYESALNDSFWGY 6 1K22 Q I 6 L 4 D V E L K G

: 140 : 95

HPFlhA : HPFliA :

* 160 * 180 * 200 *IRSVLQALLHEKIPIKDMLTILETITDIAPLVQNDVNILTEQVRARLSRVITNAFKSEDGRLKFLTFSTDAKTRVNGAMLDYLRSLDVISR-SSRKLIKSIDIEITKHLNEHGKEPSDAYLAQTLGENIEKIKEAKTASD 43 6 6 6 D663 3 I 6 L E 4 6 46K 3D

: 210 : 164

HPFlhA : HPFliA :

220 * 240 * 260 * 280SEQFLLNKLRENGTSKSLLLNVGELQKLIEVVSEEAMKVLQKGIAPVILIVEPNLRKALSNQMEQARIDVIYALVPIDEQFNAIEQDEITKKIEAEELLEHVQKALNQMSEREQILIQLYYFEELNLSEIKEILGITESR 6 N 6 E 2 L6E V 6 24 6 L L 26

: 280 : 234

HPFlhA : HPFliA :

* 300 IVLSHAELDPNSNFEALGTIHINFISQIIKEVIKKVR-KSLGVDHG--I E6 LG H

: 304 : 255

Page 3: Rust et al., Figure S1

pCJ10014603 bps

BamHI

EcoRI

flhAc

T25

pCJ10014603 bps

BamHI

EcoRI

flhAc

T25

pCJ10043239 bps

BamHIEcoRI

flgM long

T18

pCJ10043239 bps

BamHIEcoRI

flgM long

T18

A B

Rust et al., Figure S3

BACTH plasmid constructs coding for HPFlhAC-T25 (A) and HPFlgM-T18 (B) fusion proteins in plasmids pKNT25 and pUT18, respectively.

FlhAC T25 FlgM T18

pCJ1001+pUT18

pKNT25-ZIP+pUT18-ZIP

pCJ1001+pCJ1004

pKNT25+pUT18

C) Screening of transformants containing BACTH plasmids in E. coli BTH101 on LB-IPTG-x-Gal plates. pCJ1001+pCJ1004 are transformants expressing both FlhAC-T25 and FlgM-T18 fusion proteins.pCJ1001+pUT18 and pKNT25+pUT18 represent two different negative controls (see also table 3). pKNT25-ZIP+pUT18-ZIP are the positive control transformants expressing two leucin zipper domain fusions (for the latter see Karimova et al., Proc.Natl.Acad.Sci. 1998, and BACTH manual).

C


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