Ad t d di d t fAdvantages and disadvantages of the horizontal gel system of g y

gelcompany

2DGel flatbed 12.5% NF Large vs.

traditional gel systemtraditional gel system

Knut Büttner, Institute of Microbiology, Ernst-Moritz-Arndt-University of Greifswald, gy yGreifswald, Germany

2DGel flatbed 12 5% NF Large for the2DGel flatbed 12.5% NF Large for the FlatTop-Tower (gelcompany)

• Pre-cast gels on non-fluorescent film-backing• Available pre-cast gels:

– 2DGel flatbed NF 12.5% Large (200×255×0.65)– 2DGel flatbed NF 10 - 15% Large (200×255×0 65)2DGel flatbed NF 10 15% Large (200×255×0.65)

• IPG-Slot 240 mm long• Applicable for fluorescent staining (Lava Purple Flamingo)• Applicable for fluorescent staining (Lava Purple, Flamingo) • Fluorescent film-supported gels can also be stained with Coomassie Blue• Also in small format (125×255×0.65) for 2×11cm IPGs

FlatTop-Tower 4FlatTop-Tower 4

Quelle: http://www.etcelpho.com

Running conditionsRunning conditions

• Electrophoresis programme:

Step Set [V] Set Set TimeStep Set [V] Set [mA]

Set [W]

Time [min]

1 100 7 2 301 100 7 2 30

2 200 15 5 30

3 300 30 5 10

4 1500 40 40 240

• Temperature: 15 °C

Reduction and AlkylationReduction and AlkylationGreen: 15 min Equi A (with DTT), 15 min Equi B (with IAA)

Red: Only 5 min in Equi Buffer (without DTT and IAA)y q ( )

Master Gel Image: 090317_B_subt_168 WT_Exp_BMM_Flat_40W_40mA_A_LP_450V

Sample Gel Image: 090317_B_subt_168

The short running time allows to abandon reduction (with DTT) and alkylation (with IAA) of the proteins before SDS PAGEWT_Exp_BMM_Flat_40W_40mA_D_LP_noEqui_450Vand alkylation (with IAA) of the proteins before SDS-PAGE.

Influence of the electric powerInfluence of the electric power on SDS-PAGE

In 50 W gels protein spots become visible, which are superimposed Master Gel Image: 081203_B_subt_168 WT_Log_BMM_500µg_Flat_4P_50W_1_480Vby other spots in 2 W gels, or have not been stained.

Influence of the Electric PowerInfluence of the Electric Power on SDS-PAGE

50 W Gels2 W Gels 30 W Gels

Dps, YphP0YphP

DpsYphP

Dps

RplJYdaG, RplJ YdaG, RplJ YdaG, RplJRplJ RplJ

Reproducibility: IEFReproducibility: IEF(traditional gel system)

Some protein spots seem to be horizontally displaced in the 2D gel, when IPGs of different IEF runs are comparedwhen IPGs of different IEF runs are compared

Reproducibility: IEFReproducibility: IEF(FlatTop Tower 4)

Master Gel Image: 081203_B_subt_168 WT_Log_BMM_500µg_Flat_4P_50W_1_480V

Sample Gel Image: 081203_B_subt_168 WT_Log_BMM_500µg_Flat_4P_50W_2_480V

Also comparing 2D gels with IPGs from the sameIEF run shows horizontal displacements of some protein spots.IEF run shows horizontal displacements of some protein spots.

Reproducibility: SDS PAGEReproducibility: SDS PAGE(traditional gel system)

In the traditional gel system there are often serious differences i b diffappearing between different runs.

Reproducibility: SDS PAGEReproducibility: SDS PAGE FlatTop-Tower 4

Master Gel Image: 081203_B_subt_168 WT_Log_BMM_500µg_Flat_4P_50W_1_480V

Sample Gel Image: 090223_B_subt_168 WT E BMM Fl t 40W 40 A B LP 200 i 1

The commercial pre-cast gels from gelcompany offer very good reproducibility over long time periods.WT_Exp_BMM_Flat_40W_40mA_B_LP_200microns_1reproducibility over long time periods.

Staining: LavaPurple vs FlamingoStaining: LavaPurple vs. FlamingoGreen: Flamingo-Staining

Red: Lava Purple-StainingRed: Lava Purple Staining

M t G l I 090114 B bt 168 WT L BMM 500 Fl t 4P 50W B Fl 380VMaster Gel Image: 090114_B_subt_168 WT_Log_BMM_500µg_Flat_4P_50W_B_Flam_380V

Sample Gel Image: 081126_B_subt_168 WT_Log_BMM_500µg_pH 4-7_Flat_4P_50W_480VLava Purple stains acidic, low molecular weight proteins less efficient than Flamingo, high molecular weight, basic proteins are stained better with Lava Purple than withFlamingo.

Staining: Lava Purple vs FlamingoStaining: Lava Purple vs. Flamingo

Master Gel Image: 090119_B_subt_168 WT_Log_BMM_500µg_Flat_40W50mA_A_Flam_430V Sample Gel Image: 081203_B_subt_168

WT_Log_BMM_500µg_Flat_4P_50W_2_480V

Flamingo Staining Lava Purple StainingFlamingo Staining Lava Purple Staining

Flamingo often causes a black shade on the pre-cast gels.

Staining: Colloidal CoomassieStaining: Colloidal Coomassie

Sample Gel Image: 090526_NCF_10grad_1The pre-cast gels on non fluorescent film-backing (for Fluorescent staining) can also be stained with colloidal Coomassie.staining) can also be stained with colloidal Coomassie.

Identification of ProteinsIdentification of Proteins

Horizontal Gel

Identified Proteins 885

Traditional Gel

Identified Proteins 705

(without duplicates)885

Number of identified spots 1548

(without duplicates)705 

Number of identified spots 1206 

Sum of identified Proteins, which have not been

identified in the vertical gel269

Sum of identified Proteins, which have not been

identified in the horizontal gel

90 

Additional not identifiedSpots 

406Additional not identified

Spots 282 

b f i i hNumber of Proteins withexactly two Spots

178

Portion of Proteins 12 %

Number of Proteins withexactly two Spots

119 

Portion of Proteins i h l 2 S

10 %with exactly 2 Spots

12 % with exactly 2 Spots%

Proteins, which have not been identified in the Vertical Gel

Smc YfkN YfkN

the Vertical Gel

ThdF

YosN (NrdE, YosN)

YosN (NrdE, YosN)

PolA (AlaS, PolA)

LeuS (LeuS, PheT)

RecN (ProS, RecN)

YqeV (YqeV, IlvD, GcvPB)

YtcJ

YfmG

YqjN

LepA

GroEL

GabD (PdhD, GabD)TuaD

SpsB

MmsA (MmsA, YfmT)

PhoD

YdbR (YuiE, YdbR, SerA)

MalA, MenF (MalA, MenF, PdhD) YabN GltB

YerQ

YtzF

YkfA YfnD

SpsB

RsbU AcoA (Hag, AcoA)

PatA

AraR

YlxA

YkfC

YkpB

AroD, SpsKYlmD

MinC (MinC, YpjG)

YitH AcuB (YgxA, AcuB)

DnaI

OpuBA (OpuBA, PatA) YitA (Sat, YitA)

HemAT (HemAT, MurAA)

MurC

YxeP (AspB, YxeP, YvlB)PatB

Buk (Buk, AroA) YrvO (CitZ, YrvO)

YxzC (Mbl, YxzC) YoxA (BioB,YoxA)

YtqA

LacR (YfnG, LacR)

YfnE

YkkE GltC

YdhC, YtdA (YlnE, YdhC, YtdA)

YwqL (YwqL, PurF)

YhfS (YhfS, AroA)

GutB (IlvC, GutB)

YcsN PksD

HemE

YviA, DnaX

ThyA (ThyA, ResD)

YodE (FabHA, DapB, YodE)

TrxB FruK

PyrD (YaaD, PyrD, FabHA)

ThiL (MetK, ThiL)

YdbD YtbE

YitL (YitL, Hag)

YxbB

YdfO (Hag, NadE, YdfO) YvzB YbgG

GntZ (YqjI, GntZ)

( )

ProA LpdV (LpdV, ArgG) YoaC (YoaC, Obg)

YclM

Drm ArgF, YjcS (Asd, ArgF, YjcS)

Drm

YyaF (LeuB, YyaF)

PrfA (PrfA, FtsZ, Icd)

HemB (MreB, HemB)YbfQ (YwaA, YbfQ)

IspA

EtfA YhxD (YhxD)

YvzB

DapF PstS

YxaA (YdiR, YxaA)

PepT

AccC (AccC, YbbT)

PurT (TufA, PurT)

YbgE

YfmJ, YosP (YfmJ, NrdF, YosP)

Y H

EtfB YvrE (PpaC, YvrE)

SdaAA YurL (Sat, YurL)TrmD

NfrA

YhdZ

YbaJ (FabI, YbaJ)

AlsD (Sat, AlsD)

RsbX RsbX YsiA

QcrA

YxkC

SpoVFB

YknY

YusA (RpsC, YusA) NfrA YlnD (YjbV, YlnD, YkrS) EcsA

GlcR

GamA (GamA, FabL)

HisH

RibE

YpiB (YpiB, AroK, Xpt)

YheG YqgN

MsrA

PyrE YdfN

YtqB

YdaF (YdaF, YwjH)

YueJ

YvdT

YjcK

YrdA (YrdA, RpsG)YppQ

YjkB (YlmE, YjkB)

YhfI

RnhB (FabI, RnhB)YwfL

YfnH

UppS (UppS, SpsI)YqeU (SrfAD, YqeU)

YhdZ YwnB YkvJ

YgaC (LuxS, YgaC)

Hpr

YvqK

YrpE (YrpE, YwrO)

YlaL (YlaL, YwrO)

YacN (YacN YjoA)FlgC

Ki C

YsmB, PheBMoaE

YncF YosS

MoaB

YsnB

YfiR (SodA, YfiR)

YtaG

YezE (Efp, YezE)

Tmk

YsiB

DppA (MinD, DppA)

YcnD

YabD (YabD, TenA)

YsnA (YsiB, YvbH, YsnA)

PabA (PabA, HisA)

YdgI

YcnI (MreB, YcnI)

AroC (TenA, YwfI, AroC)

YfjR (Rph, YfjR)

KduI (KduI, YaaD) YycF (CodY, PurC, YycF, YurL, Mtn)

YdbD YtbE ExoA (ExoA, YwtE)

RsbR

YngH

PhoP (PhoP, YlmA)

YokJ

YwqK (YwqK, Tpx)

InfC (InfC, YpjQ, YceE, YceD, )

YpjQ

YuaC

YcsE (Adk, YcsE)

YuxI (RplC, YuxI)

RplO

NusB

YtpP

YacN (YacN, YjoA)

YpdQ YwpF YrhF

RpsI

RsbT, RpsQ, YdcD

RplV (RplV, RpsQ)

KinC

YusI

YozA (YozA, RplS)YusE

RpmA

AroH FliT

YqaE YppE

AtpC

YmcA (YqhS, YmcA)

YjbI

YunG

RplK Abh

YusU

ThiS

SpoVS

YutD

AtpF

YozE

YjcQ

FolB

YcnE

Crh (RpsS, Crh)

YpjD

YdcE

Veg, YerC

YwdI YopJ

YwqH SpsL (Nin, SpsL)

YxbD AtpF YllB

YxiF

YncF, YosS

YrrK

Spo0F

YqhY

YdbP (CspD, YdbP)

YetH, RpsP, YorD

YlxM, RpsQ (YlxM, RpsQ, RpsD)

YutI (YutI, CspD) YitW

YjbL (YjbL, YjlC)

RpsJ (YpfD, RpsJ, RpsS)

RpoE

YeeF

FhuC

YphP (Dps, YphP)RplR (RplR, RpsM)

YvbK

RplS (YwbC, RplS)

YhfF

YhcB

YdiB (YhcB, YdiB)

Def

YaaK (YaaK, Apt)

YxjI (YkrT, YxjI)

YtsP

YkvS, YqfC

RpsM

YwhB

YozE

AtpF

YlaN (YkzG, YlaN) YkuJ

YfkK YpmA

YaaA

When more than one proteins have been identified in one spot, before

Sample Gel Image: 090526_B_subt_168 WT_Exp_BMM_450µg_Flat_A_NCF_460V_100mic Created by Delta2D - http://www.decodon.com

e o e t a o e p ote s a e bee de t ed o e spot, be o ethe brackets there are the proteins, which have exclusively beenidentified in the horizontal gel, within brackets there are all proteins, which have been additionally identified in this spot.

Proteins, which have not been identified in the Horizontal Gel

CgeA

YetO (YvgR, YetO, YkfD)

YvbJ

PnbA (PnbA, PdhD)PbpEB lH (B lH Y T)

YpwA

YheB (AspS, SrfAB, SrfAA, YheB,)

YmfA

SpoIVABkdBYcgN

XynB

YhcX (ThiC, YhcX)

YusJ (YitJ, YusJ)

YdiF

MutS

RocA (LeuA, RocA)

MenD

YocI PucDYokI (YokI, DhbE)

KatE

LonA

LonA

YsnF

FliYSpeA (SpeA, YfmT, YueK)

FliDPbpE

FtsA

PbpE

GlpKYrrO (YrrO, GatB)

BglH (BglH, YugT)NadB (AspS, NadB)

DnaC

YqbM (FabF, YqbM)

DacA (YqjI, DacA)

AlaT (HisZ, AlaT)

RapGYugK

DnaN (SucC, DnaN)RbsK (NusA, RbsK)

SpsEDxr (Dxr, YbaL)

PurK (PurK, YqiG)

ManR (IolS ManR)

OppA

CitA (YjcJ, CitA)

YacK

YaaN (YhxA, YaaN)

YloI

DegS (DegS, YydG)

YxeQ

MelA

YaaH

YpwA

LicH

AnsAYloC (Mdh, YloC)

HprK

YvrH (YvrH, TenA)

YtbQ (YtbQ, YwfI)YojG

YwqC (TufA, YwqC)YwfCYfiY

YcdF

GlcK (GlcK, RpsB)

YfmF

FabHB

YabC

YvcK (SucD, YvcK, YfnG)

YclP

SpeE Gcp

ManR (IolS, ManR)

CssRYtkL

( , )

YqiX HxlA

YdfQ

YqeK (AroK, YqeK)HisB (YwjH, HisB)

YcgM

YkrY

ClpQ (YwiB, ClpQ)

GsiB RplM

RpsE (RpsG, RpsE)

YsmA

YwkD

When more than one proteins have been identified in one spot, beforee o e t a o e p ote s a e bee de t ed o e spot, be o ethe brackets there are the proteins, which have exclusively beenidentified in the vertical gel, within brackets there are all proteins, which have been additionally identified in this spot.

Mastergel Flat Top Tower (Horizontal Gel)

Smc SrfAC SrfAC

PycA PycA

SrfAA

PycA SrfAA,SrfAB

PyrAB

YfkN

SrfAC

GltAGltAGltA

PycA PycA

SrfAA

SrfAB SrfAA,SrfAB

PyrAB

YfkNRpoB RpoB

YlaG

DnaK,YvgR

YuiE

YurU

HtpG HtpG

Hag

FusA

FusA

PurL PurL PurL

FusA

SrfAB,SrfAA

FusA

LigAInfB

MetE

MetE

LeuS

AroA

IlvB

YjbG

YlaG

AtpAMetK

AtpA

DhaS,YfmT,YueK

Hom

PdhC

GatBYurX

YurU

PdhC,YurU

PdhD

GuaA,PdhD

AlaS,PolAAlaS

YlaG

YlaGMetS

YlaGGlyS

PnpAMetS

Vpr

YitJ

IlvB FabHA

GlyS

DltAYurU

YkpA,LysSGltX

PurHAsnS

SerSPabB

AsnS,YueK

SpsB

MmsA,YfmT

ThiC

Zwf

YqeV,IlvD,GcvPB

ThrS

IlvB IlvB

ArgS

YqjNPgm,PckAPgm

IlvD

ThiC

ThrSThrS

OdhA OdhA

Ald

AldIleSIleS

IleS

Pps

SecASecASecASecA

PrkA

GyrB

SdhA

ThiC,YitJ

ThiC

PrkA

ThiC

LeuA

LeuA

SerA

YvgQ

SdhA

AcsA

YvgQ

YtcJ

SerA

LeuA

PurF

MurD,YvfW

MurDLeuCFliI

AroF

AroE,TufAAroE

GuaB

GuaB

GuaB

KatA,YtcI

GuaBGuaB

KatA

YhfT YvfW

YkqC

YvfW

SerA,FabL

PurF

TuaD

SerAGcaD

SerAYvgQ

DhbE

AsnBAsnB

DhbESdhA,YkqC

ThrZ ThrZ

OdhAOdhA

RpoC

ClpCClpC

NrdE,YosNNrdE,YosN

GidAGidA

ThrS

YjbG

AtpDAtpD

GroEL

GroEL GroEL

GroEL

GroEL

GroEL

PtsI

DnaKPtsI

DnaK,YvgR DnaK

SrfAASrfAA,SrfAB

CarB

CarB

CarB

CitB

PyrABPyrAB

PyrAB

RpoBSrfAC

CitBCitBCitB

AlaS

ValS ValS

PheT,LeuS

MetEMetEMetE

MetEMetE

ProS

PnpAPnpA

LeuS,PheT

MetE

AhpFAhpF,SigA

ThdFSigA,YtjP

AspSAspS

GlmSLepA

AspS

TktTkt

Pyk,YhxBPyk

ProS,RecNMetE

ProS

GlmS,AdeC

GlmS

Hom,PdhCPdhD

HisD

OdhB,GatBGatA

YopQ

PurHCysS,PurH

SpsB

PurH PurH,YueK

LysS,GltX

Pgm

YfmG

YllA

PyrGPyrG

ThiCYfmR

ThrS,Fbp

ThiC

YshC

GcvPB

IlvD

SdhA

IlvC

ClpY

GcaD

PurA MurF

PurA

CitGMurD

DnaA

PurF

DhbE

PcrA

DhbE

PurF

YvfWYvgQ

PurA, MurF, CitG

YkpA, SrfAA

PurHYurX

ValS

LigAMetE, PnpA

AspS, DnaK

YlaG

PnpA

AtpD

GroELTig

GidA

NrdE

TagE

DhbE

IleS, SrfAB

SrfAASrfAA

RpoB

RpoB, YrrC

SrfAA

SrfAA

PyrG Pyk

YlaG, DltA, YurU

YlaG

Zwf, AsnS

LicHGlyA, CitG AtpA

PnpA

SrfAA

HtpGYvgR

ValSSrfAA, SrfAB

SrfAB, SrfAA

SrfAB, SrfAA

SrfAA, SrfAB

AtpD

AtpD

GroEL

GltB

YpiA,AroF

Tig

TigYloV

PhoD

GltB,FlgKGroEL,YdbR

YuiE,YdbR,SerA

FusAYabN

GroEL,DnaK

GroEL

PtsI, GroEL

DnaKThiC, DnaK

SrfAA, SrfAB

CheA

PurL

PurL

SrfAB, SrfAA

PurL

SrfAA,SrfAB

SrfAB, SrfAA

SrfAB, SrfAA, PurL, FusA

FusA

FusA

DhbF

PyrAB

DhbF

SrfAC

FusAFusA

MetE

PdhC

PdhD

PdhC

PdhCSrfAA MetE

GlmS

Pyk,YhxB

Tkt,YhxB

Pyk

ClpXHisD,ClpX

PdhD

PdhD,GabD PdhD

GuaA

AhpF, ThdFMalA,MenF,PdhD

BglA,PdhD,GuaA

SrfAA, SrfAB, DnaK, AspS,

DnaK

DhaS,AtpA,YurX

YurX

GlySGlyS

YlaG

YjbG

PyrG

ThrS

ThiC

YitJ,PyrG

YkoD,YdaP

P B

GidAroF

AroF,YhfE

PurF

OdhB, GidPurA, MurD

AnsB,GcaDLeuA

GntK, SerA

ThiC

ThiC, IlvC

YitJ,ThiC

ThiC

CitZ,NrdE

YkqC

GuaB, LeuC

LeuC

GuaB

GidA

YueK,CysS

YokG

YfnF

YxbBTufA

FabD,PdhB

RpoAGroEL

GroEL

Eno,YkuI

YciCYciC

DhbC

TyrS,DhbC

ProAYqjI

GatA

AtpA

MleA

GlnA,MleA

LysA

YtsJ

DrmPrfA,FtsZ,Icd

SucC YqhT YhdN,SucC,GapA

GlyQYojN,Dat

BkdAB

Mdh

BkdAB,FusA

YkrS

YkgB,YvaA

HemL

AckA,DhbC,YvaA

PheS,Ald

Csd

HisS

HisS

YqjI,GntZ

SerS

YkvY

YhfS,AroA

YulF

YumC

YkrSYxjH,Hag

YxjG,YhdR,YkrT

RecAYkrS,SucCHisC

YwaA

YbgE

YvzB

YaaD

SigBYqfS

IlvC,YoaD

GlyAHemAT,MurAA

YhfE,AroF

PurH

CitZ

CitZ,YsdC,SerC

IlvC,YvlB

YphC

BioA

YcgT

ThyA,ResD

SucD

YqfL

FolDAccD

SerAGsaB

RibA,YhxAGudB

NadA

CggR

Prs

YyaA

AccD,RpsB,CysK

PheA,PurRLeuA

AccD,SpoVFAAroD,SpsK

RpsB

YkpB

OpuCA

YjlD PdhAGcvT

YjlD

PdhA

LeuC

PfkAAccA PhoH

GuaC

YlxA

MetA

YpdA,PfkA

YutJ

BioIYqeH,BioI,Zwf

YqeH

BioI

PurD,TufA

AccA

YjcH

YvfV

YlmD

YqhM

EtfB

PstS

Hag

YvzB

YdjE

PpaC

FliGPdhB,TufA

PdhB,TufAPta

EtfA

Mdh,Hag

PanC

HemH,Pta

NusA

TufA,PdhB

TufA

IspAPurM,RpoA

YxbCGroEL

PurD,LysC,TufATufA

YbbTFusA

YpfD

Eno

YbbT

YtjPYbbT

AccC,YbbT

ClpX,ThdF

Obg

HisD

ArgH

YqjE

InfB

Pgi

TufA

TufATufA

LeuB

LeuB,YyaF

Pgk

Icd

YdjL,SucC

TufA,PurT

LpdV,ArgG

YlmB,AmpS

LysA

YaaDTrxB

MreB,HemB

NadE

YkrT,YkrV

YkrTYjcI

Bcd

YhdR

YxjG

FtsZ

YdjL

Drm

NusA

LeuB

SucC

YxjG,HisC,YxjH

GapA

YwaA,YbfQ

Hag,AcoA

Hag,YvzBTsfHag,NadE,YdfO

RsbU

BkdAA

SucC

TufA

GalEGalE

YfmJ,NrdF,YosP

Hag,YvzB

YtqI,YhfPHag

Hag

YbgG

Hag,FusA,Mdh

YcdH

FruK

Tsf

Tsf

FbaA

GtaB

Dat

Tsf

Tsf

GlyQ,DapA

Mdh,Icd

ThrB

YitL,Hag

SpeBYacC

YdbD YtbE

NadC

YjjAYsgA

FbaAFbaA,YvgN

YaaD

FusA

MenB

MenB

NadC,YjjA

Y fL

MenB,FolDMenB,YtpQ

RapF

RecA,ThrC

YcsN

YqkF

CcpA

Tsf,YaaD

MetK,QueAYvcT

ArgC

GapA

AroA,GapA

Sat

YugJ

AckA,AldHisZ

YjcI

Asd,ArgF,YjcS

YtsJ

MetK,PyrC

DhbC

PurH,DhbC

PyrC

OpuAA,HisS

YqiG

DhbC

MlpA

HisS ArgG,YqjI

MurC

BioA

PurA,MurF

GlyA

CinA,Sat

AspB

YwfGAspB

AspB,YxeP,YvlB

PatB

AspB

YkrW

YkrW,SatAspB

Sat,YhaA

YhaA,GlyA,Sat

OdhB

YvyHNadA

Sat,YitA

GsaB

SatSat

IlvA

IlvA,GlyAYxeKYhxA

GudB,GudB1

MoeA,AmhX

YsdC,GlyA

PdhA

YjcJ

PyrAA

FolD,YtzF

AtpG

SucD

Buk,AroA

IlvC

BioB,IlvC

PyrB,BioB

YeaC

BioB,YoxA

PlsX

YtoPYtoP SerC

YsdC

YsdC

CitZ,YrvO

IlvC,Cah,YkcA

IlvC,TyrA

IlvC

AroA

YurP

IolSIolS

YfnE

YurP,Mbl

YtqAYfnG,LacR

RpoC

YfnDYbbI

Fmt,PyrB

YeaCYfnG

ThiF,MblMbl,YxzC

Fmt

DnaI

ExuRYqfP,YhaM

YocD

SerA

FabI

YusV

PurH

Sul

YerQ

YkkEGltC

PurR

PurRPdhD

YtzF

CysK

FabI

PyrK PyrK

YflG

FabI,RnhB

Yi SYwqL,PurFYcbG

YtrE

YvyE

Spo0A

ArgB

CwlC

YjbN YlnE

MetK,SpsD

YitH,YlnE

YgxA,AcuB

YlnE,YdhC,YtdA

RpsB,AccDSpsD

RpsB

KsgA

Prs

YfhM

RpsB

RpsB

PfkA,AccA

PrsYyaA

YhaM

GuaC,YqjK

YhfJ

Tgt,YutJ

ArgD

YjlD

GoxB,YutJ

PatAOppD

GoxB,MurAB

DapGYjlD

YutJ

YjlDAraR

YjlD

SerA

DapG

OpuBA, PatA

OppD

YjcJ

Mbl

YceH

Prs, LeuC

YlnE

YvfVYitH

PheA

LeuA, PheA FabI

IlvC, YjlD, IolS

SucD, YqfLHag, Tsf

Csd

DhbF, YdjL

NusASucC, YbfQYkrS, GapA

Mdh

MetE

PpaC

MetK

PrsCcpC

YhfJ

YjlD

SerC

GlyA, CitG AtpA

NadC, YsgA

YwqG

YokG

YfmS

RpsBSpo0M

MreB,YwaA

PpaC,YvrECheV

YwqG

YceD, YceE

YceD, YceETufA

YhxD

DhbB

AcoB

DhbB

YdiR,YxaA

PurM

PurM

PurM

RpoA

RpoA

PepT

Eno

Eno

FusA, YpfD

LeuB

YxjG,YxjH

PurD

YpfD

FusA

YpfD

YclMOdhB,ArgH

ArgH,YmfH

GlnA

FabF

YmfH

YdjL,YkrT

ClpX

Pgk,Icd

YqjE

SpeB

YngH YkrT

DapF

Hag,YvzB

Spo0M

HagFlgL

ThiD,DapA,RocF

Tsf,YaaD

FabHA,DapB,YodE

YaaD,PyrD,FabHA

Tsf

FtsY

FtsY

YwaA

IlvC,GutBAroA

YbaL

YxjG

DhbC

YoaC,Obg

Pgi,MetK

GlnA

Pgi

PrfB,GlnA

YwaAYumC

YumC

YdjIMetK, YvcT

YwjI

YwjI

AroF, ThiC

PdhA

YdbM

PdhA, YdbM

YjcJ

PyrAA

YceH

PurB

ArgD

Prs

MetA,AroA,YlxAPrs

CcpCRpsB,CcpC

YkfC

MinC,YpjG

RpsB

RpsB

MurBYhaQ

PfkA

GpsA

YisK

YkfA

YkrS

IlvC,YtxK

YurP

MetK

FolD

CysK, YfjP

HemE IlvC,IolS

PksD

LeuA,MenB,DhbE

NadCYfhB,YqiD

YviA,DnaX

MenB

MenB,YtpQCysK

YrdA,RpsG

TrmD

PhoP,YlmA

YwjH,YodD,YueJ

YciC

YsaA,GtaB

AccB,RpsD

MreB,YcnI

Efp

NudF

TufA

KipI

RsbR

YwtE

TasACodY,PurC,YycF,YurL,Mtn

YshC,YvaK

Tmk

PabA,HisA

ComA

RplC,ComA

RplC,YwfI,TenAYabD,TenATenA

YsiB,YvbH,YsnAYsiB

LysC,YkaAYhfK

TufA

YocJ,YrdC

YvdD

MoaB

YtxH

YncF,YosS

YutG

DivIVA

Ssb

YkvM

MoaE

YpqE

YwrORimM

YkvJ

YcsF

YjbV

EcsA

PyrE

YwjH

BsaA

BsaA,YkrW

AroA

SerA

YwiB

YqzC

YwpBYppQ

YsiA

GrpE

ArgJ,YceD

AhpC

InfC,AhpCYokJ

YkrZ

RsbW

YqaPYwqK,TpxTpx

AptRpsB Tig

Tig

ClpY

YdgI

YuaC

HprT

GrpE

YmaB

YkuU,YkrTLeuD

YhcW,YkrX

Gmk,GtaB

ThiG,YurY

PurQ

YlbN

RpoA

YxeH,Sat

Sat,AlsDTufA,YxeH

GtaBYaaD,TrxB GtaB

Gmk,RpsC

YurY

YkoBSdaAA

Sat,YurL

YkoB

YkrT

TpiA

FlgETpiA

PunA

CodY,PurC

PurC

PurC

Rph,YfjR

KduI,YaaD

YurY

TpiA

ThiG,YkuQ

YkuQTufA

YqgX

TufA

TufA

TenA,YwfI,AroC

Soj

TenA

HisA

DeoD

YkaA

SodA,YfiR

Efp

SodA,YaaE

RpsB

PyrR

YaaE

Efp,YezE

YtaG

YkrT

DivIVA ClpP

YwnHYsnB

TufA RplJ

YlmFYuaI

DfrA

ClpP YocJ

TufA,LuxS

HemL

MinD,DppA

YcnDYktC

FolD

ThiE

PyrRFusA

CysC

CysC YvaB

YodC,AroAYkrT

YodC,AroA

NusG

YvqK

YrpE,YwrO

PdhA

SrfAD,LexA

SigH

LeuA

Hpr

RpsD,BioW

YhfRYvyD

YwnB

FolD

HisFYkwC

YvbY

PanB

SrfAD,YqeU

DhbADhbA,PdhA,CysH

HisF

YddKYddK

SigD

YjjA

MenB

PyrH

YsgASerA,GapAYwfLFabI

SpsA

UppS,SpsIDhbA

AroA

PksC,FabIArgG

YfnH

ResDCysH

YhfI

SucD,RplC

YitSGlcRYjbV,YlnD,YkrS

NfrANfrA

YcsF

DegU

DegU

PspA

GamA,FabLPyrF

YfkO

YodC

FrrYoxD

YceCYlpC,PurB Udk

YceC,AroA

RpsD

Pyk

YlpCRpsDYkrB

YdiC,YceCHisH

OdhB

Maf

YheG

YgaF

MsrAYwjH

SerA

YtqB

YwjHYueJ

YwjHYwjH

LuxS

YdaF,YwjH YjcK

YjcG

YrkLTufA

YkrT

SerA,YgaF

LeuB

YjcF

FlgCY N Yj A

RplK,YsmB

YkrT

YtkDYgaF

YuaE

YtiB

RibEUpp

YrxA

PurN

Xpt

AroK

SpoVFB RpsD,AroA

YjbKRpsC,YusA

RsbX

YktBYkoF

YbxB

FabL

YufM

YqiKYknY

DhbA

YpiB, AroK, Xpt

RsbX

YvdT YhfK

Frr

YoxD

YcdH

SodA, YaaE

ClpP

TenA

YwfI, TenA

YktC

FabI, YbaJ

RpsD

TufA, YfkM

YraA,Ssb

YraA

YesJ, DivIVA DivIVATpx

YceE, YceD

FabL

Yj A

YlbI

YjcG

PurN

DhbAYfnHFabI, YqfN

YqeU YkwC HisF

YwfC

TenA, YvrH

YsnA

YdjL

DivIVA

RpsG YokL, RpsG

TenA

TufA

HprT, InfC

YkrZ

GreA

InfC, YpjQ, YceE, YceD,

CheW,InfC

LysC,CheW

YkrZ

YceD,YceE,AhpCAhpC

YtpR,YceD,RpsD,YceE

TigRpsC

AccB,YceE,YceD

YceE,YceD

YpjQ

LeuD,RpsD

MreB,FlgE

AccB

Adk,YcsE

HisG

TufA

YkrA,HisG,HisI

HisG

YutF

YsaA,HisI

YutF

TufA

TufA

TufA

TufAGtaB Ctc

YeeIYeeITig

RpsB

Eno

RplC,YuxI

RplC, RpsC

YfmS,YdjL

YfmS

RpoA,YfmS

ExoA,YwtEThrB TufA

YwjBSsb

YraA,GroEL

LysC

SodA,YaaE

RpsD, TufADra,GtaB

TufA

RpsGRplJ

YhfK

LuxS,YgaC

YoxD

YvyD

Pyk, YhfR, SigH

YvbYYhdZ

YhdZYwfC

YvaK

YvaK, YkrUYkrU,FbaA

YaaD,TrxB

Cmk

PyrR

CysC, CmkPyrH

YwjH

YlaL,YwrO

TufA YtoQ,TufA

YtoQ,RpsG

FurBsaA

YtoQ

YjcGYqgNAroK

YkqB,Rpe

Dgk, RpsD, Upp

Rpe,Pgk

SpoVC

YdfN Dck,YdiC Pgk

DegU,YlmE

YlmE,YjkB

YitV

DegU

NagB,YjdA

YdiC,LeuA

Upp,RpsD

RpsD, Pgk, HisH, Upp

QcrA

CspR

RpsG

YxkC

YaaQ Hit

KinC

YdfG

RpsGYqaP

YuaE

BioD

YtmQ

YeeF

RpoE

TufA YllB

YhgC

YrzC

YhcB,YdiB

YncF,YosS

Def

RpsG

Nin,SpsL

IlvC

YwqHYurV

AhrC

PurE

RibH

Ndk

PurH

RsbT,RpsQ,YdcD

RplV,RpsQ

YkrZ

YtpP

YpfD,RpsJ,RpsS YukE

YutI,CspD

YutI

YitW

TrxA

YetH,RpsP,YorD

YkzG,YlaN GroES

YjlC

PurS

PurS

Spo0F

YlxM,RpsQ,RpsD

CspD,YdbP

RsbV

TrxA

YheA

CspD,RplL

YhfFYqhY

RplR,RpsM

YwbC,RplS

Dps

YutE

YvbK

YlxSRplO

YaaK,Apt

YkvM

AlaR,YkvM

YslB,YqiWYqiW

FliSMgsA

YwdI YkzAYopJ

NusB

Veg,YerC

YdcE

YpjD

YlbA

RpsS,Crh

RplR YcnE RpsF

PtsH

CheY

GtaB

Icd

RpsD

YutD

MetKAtpFAbrB

YunG

YvyD

YjbIYqhS,YmcAAtpC

FolB

FliS

YppE

PurE

YugI,RplS

YusI

TufA,YwiB

YacN,YjoA

YjoA

PpiB

AroH

FliT

TufA

AroA

YrrKYqaE

YpsB

YugI

YjcQAroA

AbrB

YozA,RplS

YusE

AroA

YrhF YwpF

YjlC

SpoVS

RplU

AbrB

SpoVG

TufA

YsmB,PheBRpsI

DnaK

YozE

SpoIIAA

PpiB

AtpF

YcsD

YabJ

YueH

AbrB

Ndk MetS, PnpA

YjoA

YxnB

GsiBYfmP

RplV

YkrZ

RsbV

YjbL,YjlC

RpsS,RplL

YjlC

RplL

YkrT,YviF

YviF,YxxG

Dps,YphP

Dps,YkrZ

YtsP

YkrT,YxjI

RpsG, IleS

YxbDYhcB

RpsF

SpoVG

RpsS,SpoVGYabJ

AroA

YdcN,YqhS

YklA

AroA

RibH,YdeM

GroEL

YvcI

YwlF

YxiF

YdaG

ThiSYloH

YusU

Abh

RplK

YneT

AroA

IlvC

YpdQYqeY Ndk RibT

RibT, Ndk

TagD

YabA

MntR

MntR,YgxA

YjoA

YjoA,PpiB

YwlF

PpiB, YwlF

YugI

KinC

PurH CspB

YaaA

YpmA

YfkK

RplD CspCYkuJ

CspBRpoA

YozE

RpsM

YwhB

YkvS,YqfC

CspC

AtpF

RplD

Master Gel Image: 090526_B_subt_168 WT_Exp_BMM_450µg_Flat_A_NCF_460V_100mic

Created by Delta2D - http://www.decodon.com

Pyk RpoBRpoB

TufA

SrfAA SrfABSrfAA, SrfAB

SrfAA, SrfABRpoB, GltA, SrfAC

PykMetE

MetESrfACSrfAC

SrfAC

SrfAA

Ald, DhbCGltAGltAGltA

Master Gel Traditional System (Vertical Gel)

PdhC

MetE, GlmS

GlmS

AlaS

MetEPnpA

MetE

Asd

ProS ProS

Vpr

YhxB

YlaGMetE, YhxB

TktTktTkt ,PykPnpA, YdiF

PnpA PnpA

YdiF

MetE

MetE

MetE

MetE

MetE

PyrAB

PheTPheT

PheT ,ValS

Pyk

PheT, PyrAB

RpoB

RpoB

RpoB

SrfAB, SrfAA

SrfAB, SrfAASrfAB, SrfAA

SrfAA, SrfAB

SrfAA, SrfAB

SrfAB, SrfAA

FusA FusA

CgeA

Tig

GlyQ

CheAPurL

PurL, SrfAA, SrfAB

PurLPurL

SrfAB, SrfAAFusA

FusA

SrfAA, SrfAB

DnaK

DnaK, CheA

SrfAA, SrfAB

DnaK

AspS

InfBYuiE

GroELSrfAA, SrfAB

PtsI

GroEL

Tig

YloV

ProS

MetE

MetSYlaG

MetE

MetE, PdhC

PheT, ValS

MetEMetE

MetE

MetE

AlaS

CitB

CitB

MetEValS

LeuB

RpoB

Pyk

Pps, SecA

SrfAA

SrfAB, SrfAA

SrfAB, SrfAA

YlaG, MetS

YlaG, MetS

GlyS

MetEPyk

PykPheTYfmR

YhxB

YhfKMetE

LysS

Pgm, PyrGIlvB

Pgm

PgmPyrG, Pgm

Pgm

Pgm, ThiC

YkoD, PyrG

YdaPIlvB

ThiC

AsnB

SdhA

YshCThiC

ThiC, YhcX

ThrS ThrS ThrS

DhbC

YitJ, YusJ

ThiC

YitJ

ThiC, YitJ

YitJ

Fbp, ThiC

ThiC

YitJPrkA

ThiC

SrfAB, AhpC

GlySYlaG, SrfAB GlyS

YlaG

SrfAB, YlaG

SdhA

PycA

PycAOdhA

SecA, AroA SrfAA

InfB

SrfAA, SrfAB

SrfABSrfAB

SrfAB

CitBSrfABIleS

SrfAA, SrfAB

SrfAA

PycA

OdhAOdhAOdhAOdhA

OdhA

GyrB

GlyA

SecA

GltA

FusA

LonA

LonA

ClpCClpCClpC

PcrAPcrA

NrdE NrdE PrkA

ThrZ

SerA

MenD

SerA

AsnBSdhA

SerA, AcsA

GidA

PucD

RpoC

KatENrdE

TagE

GidA

RpoB

SrfAB, SrfAA

MetE, Pyk

IlvB

Tig

SrfAA

AspS, SrfAB, SrfAA, YheB,

SrfAB

IleS IleS

MutS

PycA, GltA

DhbE

NrdE

YokI, DhbE

ThrZ

PycA

RpoC

SerA

F A

DnaK

FusASrfAC, FusADhbF

DhbEYocI

MetE

PyrAB

PrkA

LigA

HtpGHtpG

FusA

FusACarB

PycA

AlaS

PrkA

ThiC

SrfAA

Pyk Pyk

PheT

MetE

YvgR, YetO, YkfD

YvgR, YetO

GroELAspS

AspS

GlmS

GlmS

AdeC

HtpG

LigA

LigA

FusA

FusA

CarB

PyrAB

SecA

SecA

GyrB

YjbG, Pgm

DhbE

SdhA AspS

SrfAA, SrfAB

LigAMetS

YvbJ

SdhA

YqjE

PgiPgi

YlmB

PrfB

PrfB

Asd ,RecA

RapG

YkrT, YjcI, SucCYqhT

ClpX, YqhT

IcdIcdYkrT

YdjLFtsZ

YdjLPgk Pgk Pgk, LeuB

FusALeuB

TufA ,YbbT

YmfH, GlnA

YmfH, GlnAGlnA

ArgHObg

MleA

GlnA

GatB GatBHisD, TufA

TufA

YpwA

SigA

BglAYpwA

GuaA, PdhD

PurN

FliD

PnbA, PdhD

PbpE

PdhD, Hom

PdhD

PdhD

PdhC

YurX

PdhC

YurU ,YkpA

YurUInfB

Tig

FliY

YsnF

YpiA

FlgK

YciC

AtpDAtpDAtpD

OdhB, FusA FusA

YciC

YciC

DnaC

TufAYpfD

YkuI

EnoEno

RpoARpoARpoA

YxbC

YugK

GroEL, TufA

TufA

GltX

LysS, ArgS

LysSPdhC, OdhB

Hom

YurU

YurX

PbpE

YrrO, GatBGlpK

AtpA

MetK, AtpA

FtsA

AmpSArgG

AtpA AtpA

AtpA

SerS

YqjI

MetK, YqjI, LysS MetK

HomDltA

DhaS

BglH, YugT

PurH

DhaS, PurH, SpeAPabBOppA

GcaD

GcaD, YtcI

SerS

GatA

LicH

OpuAA

YqjI

YqjI

DhbC, Ald

PyrC, DhbCPyrC, DhbC, Ald PurH

ArgG

YqjI

YlmB

LysARecA

LysAYtsJ

YtsJ, DegU, AsdTyrS, DhbC, YvaA, GlnA, PheS

YkgB, YvaA

AckA YxjH

YxjG

YkrV

YkrTYxjGBcd

HisC

YhdR, YjcI

YkrS

YjcI

YxjG, HemL

TyrS, PheS

GapA GapA

HisZ, AlaT

YkrS

SpsEYkrS, YugJ

YkrW

AspB

CitZ ,YwfGSerC

CitZ

YvyH

YvlBYkvY

PurK, YqiGYqiG

HisS, Csd

Dxr, YbaL

MlpA, AspB

Sat, PurH

YvlB

AspB

CinA

AspB

GlyA, CitG

GlyA

YkrW

AspB

MurAA, GlyA, YhaA

IlvA

BioA, HisSGlyAGlyA, BioA

SerACsd

YfmTGatA

Zwf

Zwf, GcaD

GcaD, MurFYphC

MurF, GcaD, PurA

ZwfLeuA

IlvD

IlvDBkdB PurH

AsnS

PurH

YueK

SpeA, YfmT, YueK

CysSPckA, ArgS

CysSArgSIlvDIlvD

YcgN

GcvPB

XynB

SerA, YvgQYvgQ, SerAYvgQ

LeuA, RocA

SerA

YvgQ

SerASerA, YkqC

ClpY

GntKLeuALeuA

GcaD

PurF

PurF

YvfW

NadA

YaaH PurA PurA

PurA

CitG

GlyA

MurD

CitG, NadA

RibA, AmhXOdhB

NadAYxeK

GudB, GudB1

YhxA, YaaN

YhxA

PurBDapG, PurB

MurAB

PurB

OppD

GudB

MoeA

GsaBYsdC

YqeH

Tgt YloI

YutJ

BioI

YutJ

MelA

YhfE

MurAB

AroE FliIAroF

GoxB

LeuC

SerA

LeuCLeuC LeuC

GuaBGuaB

DnaAGuaB

PurF

PurF, GuaB

GuaB

YtcI

YkqCPgi

YciC

YmfA

AspS, NadB

AtpD, GroEL

AhpF

AhpF

AhpF

PurDPurD

YbbT, TufA

TufA

TufATufA

TufA

YtjP

TufA

AspS, TufA

PurL

YlmB

YqjI

InfB, YqjE

YqjI, DacA

OdhB

GatA, YopQ

YvfW

RpoC, GsaB

OpuCA DegS, YydG

Tgt

YxeQ

Gid

SerA

LeuB, LysC

AtpD, YbbT

SpoIVA

FusA

IlvB

FabF, YqbM

GroEL

LeuC

GuaB

YhfT

YkqCKatA

PurF

SerA

LeuA

GcaD

DapG

YpfDYpfD

Eno

EnoIcd

GlnA, MetK

TufA

PurD

HisD

GuaA, PdhDYkpA

GltX

IlvBYllA

Zwf

IlvA

Sat, GlyA YkrW

BioA

AnsB

Sat

Sat, NadA

CitZ

YceH

Gid

YkqC

YllA

IlvD

ThrB

YacC

GlyQ

GlyQ SpeB

YkrT

CheV, TufA

YfnF

Mdh, YloC

Mdh

Mdh

FusA

Mdh

BkdABDat

YojN

Pgk

Dat

NadE

YcdH

Hag Hag, TufA

HagGalE

YtqI

Hag

YhdN

NrdF

Hag, YtqI

BkdAA YqhT

SucC, DnaNSucC

NusANusA, RbsK

HemH

PurM TufA

YdiR

DhbB

DhbB

AcoB

PdhB

DhbB ,FliG

YwqG

YokG

YeeICheV, Pta

Pta

PpaC, OpuAA

PpaC

AnsA

PpaC

PpaC, TufA

YdjL

YfmS

PurQ TufA

TufA

TufA

YlbN

Ctc

SucC

Hag

YwaA

YwaA

SucC, YkrSYkrS

YumC

YkrS

ThrC, ArgC

GapA

YwaA, MreB

YwaA, Gcp

HprKYdjI, Tsf

FabHA

DapB

NadE

Gcp

Tsf

Tsf

DapA, Tsf

YaaD

YaaD

Tsf Mtn

Mtn

GtaBYusV

MenB, YtpQMenB

MenB, SigD, YkrU

YqiD, NadC

YfhB, NadC

YfhB, NadC

YclP

NadC

SerA

YkrU

YvgN

FbaA

YaaD

NadC

TyrA, IlvC

IolS, ManR

YkcA ,IolS

Cah

YqfS

IolS

YqkF

YvcT

GapA

MetK, QueA

MetKYulFFtsY

CcpA

ThrC, RapF

FtsY

ThrC

ThrC, RapF

YugJ, AroA

IlvC

YoaDTyrA

IlvC

IlvC

YurP

IlvC

AroAYurP, IlvC

SerC

YsdC

AroA, IlvC

IlvC

PyrB

YeaC

ThiF

BioB

IlvC

PlsX

YtoP

YacKIcd

YdbM

PdhA

YjcJ

YjcJ, CitA

GltA

PyrAAYjlD

PdhA

ArgD

YjlD

YyaA

YhaMYhfJ

GuaC, YqjK

ExuR

YqfP

FabHB

YocD ,YhfJ

Mbl

TyrA, YtxK

YbbI

SigB

YisK

SucD, YvcK, YfnGBioW

SucD

SucD

YfmF

YflG

ResD

YurP

PheA

RpsB

YqhM

AtpG

CcpC

SpoVFA, Prs

YhaQ, RpsB

RpsB

Prs

RpsB

AccA

PfkA

YpdA

MetA

YyaA

GcvT

YjlDYjlD

PhoH

ArgD, YjlD

ArgDYqeH

, qj

Tsf, GapA

YtqI, YhfP, NusA, Hag PanC

Hag, PpaCYwaA

NadCFbaA

GlcK, RpsB

YcgT

YabCGpsA FabDFabD

KsgA ThiD, RocF

RpsBPrs

RpsB

PdhB

LeuB

YdjE

YfmS

Pta

Spo0M

YfmS

YsgA

YwjI

NadC

SpeE

CysK

Prs

AccAGuaC, PfkA

Fmt

CggR

NudF

YkuQ, TufA

Gmk

TufAYkuQ YsaA, YkuQ

YurYThiG

YojG

KipI

YwtETpiA, YkoB

TpiAYutF

TufATufA

Ctc

GrpE

Tig

RplC

ArgJ

GrpEAdk

YmaBYkrT

YsaA

TufA, YwqC

YkrA

HisG

HisI

YsaA

YkrX

LeuD

YhcW

ClpY

HxlA GrpE

YkuU

HprT ArgGAccB

YceD, YceE, RpsC

YceD, YceE

YtpR

YdfQ

Soj TasA

CodYPurC

Rph

TenA

Nap

YwfIYtbQ, YwfI

TenA, YlmA

YjjA

YvaK MinD Tsf

TenA

YkwC

FolD

YkwC

YwfC

YhfKYvbY

SigH, PanB

LexA

YqiX

FlgE, ComAPunA

YvrH, TenA

YtkL

CmkCssR

ThiE

PyrH

YktC

YcgM

RpsD

YqgXDra

HisA

YvbH

TufAYkaAYkaA

DeoD

ClpP

YocJ

SodA, YaaE SodASodA, YaaE

YrdC

PyrRCysC

YvaB

CysC, YodC

YpqEFrr

YlpC

YcsF

YjbV

Pgk

PdhA

YvyD

AroA

YhfR

IlvCLeuA

YoxD

Udk

YceC

SpsI

DhbA

CysK

AroA, PksC

MenB

MenB

SrfADYqfL, FolD

PurH YddK, YjjASul

PyrK, CysKFolD

PurR

CysHYqfL

FabI

SpsAFabI, DegU

FabI

YfiY

FabI, LeuA

YitVYjdA

PspA

OdhB

LeuA, NagB

YfkO

YkrY

PyrF

Upp

Xpt

PurN

Dck

YdfI

PyrF

Maf YdiC, Dgk

YkqB

YkqB

SpoVC, YwjH

SerA

YwjH

YkrB

YwjH, HisB

YrxA

BioD

YtmQ YbxB

Xpt

Upp

Xpt, Upp

YjbK, RpsC

YqiK

ArgB

YjcHYcbG

YpjG

YcdFYgxA

YqhM

MurBRpsB MetK, SpsD

AccD

YfhM

HisF, SrfAD, YaaD

Rpe, RpsD

RpsC

PspA

AccD

LysC

Adk

YceE, YceD

ThiG ,YxeH

YurY

TenA, PurC, CodY

YwjH

YodC

ArgG, DhbA

PurN

YkoF

YtiB

YtrE

YktB

FabL

Spo0AYufM

YvfV

YjbN

CwlC

YvyE, FabI

YlnEYutF

FlgE

FolD, YwfC

MenB

YlmE, DegU

FolD

YutGSsb

YesJ

CheW

AccB

AhpCAhpC

AhpC, YkrZYkrZ

Dps

YwjB

Apt, YitS

YlxS

GreA

Dps

Tig

RsbW

YqaP

Dps

YfkM

EfpTufA

Efp

YvdD

MreB

YraA

RplJ

YhgC

AlaRDivIVA

RplJ

YdaG

TufA, DfrA

YuaI

YhgC

NusG

YwrO

YtxH, YlmF

LuxS

RimM

YgaF SpoVC

YodD

YjcG

ArgJ

YjcG

YwiB, ClpQ

YrkL

YjcF

YaaQ

AtpD

CspR

YtkD

YgaFYuaE

BsaA

Fur

LysC

AroK, YqeK

YtiB

RplD

Tpx

YutE

YviF

YjlC

YwbCYxxG

FlgE

CspD

RplM

AlaR

YkvM

Nin

YrzC

YcsD

YurV

YdeM

YqiW

YkzAYslB

PtsH

YdcN

YvcI

YklA

FliSYneT

YwkD

RibH

YqhS

PurE

GsiB

YjcF

YtoQ

TufA

YwlF

YsmA

RibT

MntR

PpiB

YjoA

AhrC, PurE

TufA

TagD

RpoA

Hit

YugI

YqzC

YdfG

YqaP

YwpB

p

RsbV

RpsG, RpsEYwnH

YqeY Ndk

RplLCspD

YukE

CspC

CheY

PurS

RpsS ,YkzG

GroES

YheA

TrxAPurS

CspB

CspB

MgsA

YabJYtxJ

YlbA

RpsF

RplD

SpoVG

p

YugI

YpsB

YloH

RplU

SpoVG

AbrB

RsbV

Master Gel Image: 090526_B_subt_168 WT_Exp_BMM_450µg_LP_1_470V_100mic_marker

Created by Delta2D - http://www.decodon.com

Not identified Spots in Master Gel Flat Top Tower (Horizontal Gel)

406 Spots could not be identifiedMaster Gel Image: 090526_B_subt_168 WT_Exp_BMM_450µg_Flat_A_NCF_460V_100mic

Not identified Spots in Master Gel Traditional System (Vertical Gel)

Created by Delta2D - http://www.decodon.com282 Spots could not be identified

Master Gel Image: 090526_B_subt_168 WT_Exp_BMM_450µg_LP_1_470V_100mic_marker

Pros and Cons of the two Gel SystemsPros and Cons of the two Gel Systems

T di i l G l S Fl T T (H i l G l )Traditional Gel System• Running time: ca. 16 h• Reproducibility difficult

FlatTop Tower (Horizontal Gels)• Running time : ca. 5 h• Very good reproducibilityep oduc b ty d cu t

• Spots mostly homogeneous• Very labour intensive work

(casting gels cleaning)

e y good ep oduc b ty• Spots have higher conc. front• Much reduced labour intensive

(casting gels, cleaning)• Lower gel cost (excl. Labour)• Low molecular weight proteins

• Higher cost, but ready to use• Low molecular weight proteins

are often blurred, MWMin = 7.7 kDa)

in sharp spots, MWMin= 6.5 kDa)