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Table 2 Oligonucleotide primers used in this study

From: Characterization of the inner membrane protein BB0173 from Borrelia burgdorferi

Primer pair

RS

Sequence (5’➔3′)

Application

bb0173T-NdeI-F

NdeI

ACG CCA TAT GGC TTT AGC AGG TCC TTC

Amplification of bb0173 for cloning into pCR2.1 and expression vector pET23a

bb0173T-XhoI-R

XhoI

ACG CCT CGA GTA GTA TCT CTT TTA AG

bb0173cDNA-F(nt249–272)

 

GAA GAT GAT ACA TCT TAG TGC TGG

Amplification of bb0173 cDNA from RNA

bb0173cDNA-R(nt725–749)

 

CTT CCC TGA TAA AAT TTT CCA GAT

bb0173TM1-SpeI-F

SpeI

ACG CAC TAG TGG AGG ACC AGG AAA TGA GCC TTT ATA TTT G

Amplification of the first putative bb0173 transmembrane sequence in frame with the Lep construct

bb0173TM1-KpnI-R

KpnI

ACG CGG TAC CCC TCC TGG TCC CTT TAT CTT GCC TCC TCT

bb0173TM1–2-SpeI-F

SpeI

ACG CAC TAG TGG AGG ACC AGG AGA TTA TAG ATT AA TTT G

Amplification of the second putative bb0173 transmembrane sequence in frame with the Lep construct

bb0173TM2-KpnI-R

KpnI

ACG CGG TAC CCC TCC TGG TCC AAC TGA AGG ACC TGC TAA AGC

bb0173TM2b-SpeI-F

SpeI

ACG CAC TAG TGG AGG ACC AGG ACT AGA TGA TAT TTA TAT TAT G

Amplification of the third putative bb0173 transmembrane sequence in frame with the Lep construct

bb0173TM2b-KpnI-R

KpnI

AGC CGG TAC CCC TCC TGG TCC AGC CTC AGA ATG CTT TAA ATG

bb0173TM3-SpeI-F

SpeI

ACG CAC TAG TGG AGG ACC AGG AGA TAT TTA TAA AGA ATT TTT AG

Amplification of the fourth putative bb0173 transmembrane sequence in frame with the Lep construct

bb0173TM3-KpnI-R

KpnI

ACG CGG TAC CCC TCC TGG TCC CTC TTT TAA GAA AAT TTT TG

bb0173-NcoI-F.1

NcoI

ACG CCC ATG GAT GTT AAC ATT TAA TGA G

Common amplification start site for truncated insertion constructs

T7BB173

 

ATA GTA TAA TAC GAC TCA CTA TAG GAA ACC ACC ATG GGC ATG TTA ACA TTT AAT G

Amplification of BB173 with T7 promoter

TM1 NST BB173

 

TTA TCA GGA CAT CAT GGT GCT GTT ATA ATC CTT AAG TTT TAA

Amplification of BB173 56mer adding C-tag NST

TM1 QST BB173

 

TTA TCA GGA CAT CAT GGT GCT CTG ATA ATC CTT AAG TTT TAA

Amplification of BB173 56mer adding C-tag QSTMMS

TM2 NST BB173

 

TTA TCA GGA CAT CAT GGT GCT GTT AAT ATC ATC TAG CTT TT

Amplification of BB173 162mer adding C-tag NST

TM2 QST BB173

 

TTA TCA GGA CAT CAT GGT GCT CTG AAT ATC ATC TAG CTT TT

Amplification of BB173 162mer adding C-tag QSTMMS

TM3 NST BB173

 

TTA TCA GGA CAT CAT GGT GCT ATT AAG CAT ACT AGG ATC

Amplification of BB173 278mer adding C-tag NST

TM3 QST BB173

 

TTA TCA GGA CAT CAT GGT GCT CTG AAG CAT ACT AGG ATC

Amplification of BB173 278mer adding C-tag QSTMMS

TM 4 NST BB173

 

TTA TCA GGA CAT CAT GGT GCT GTT CCC GGA CAT GCT GCC CAT TAG TAT CTC TTT TAA GAA

Amplification of BB173 341-mer (full length) adding C-tag NST

TM 4 QST BB173

 

TTA TCA GGA CAT CAT GGT GCT CTG CCC GGA CAT GCT GCC CAT TAG TAT CTC TTT TAA GA

Amplification of BB173 341-mer (full length) adding C-tag QSTMMS

TM2 NST BB173

 

TTA TCA GGA CAT CAT GGT GCT GTT AAT ATC ATC TAG CTT TT

Amplification of BB173 162mer adding C-tag NST

TM2 QST BB173

 

TTA TCA GGA CAT CAT GGT GCT CTG AAT ATC ATC TAG CTT TT

Amplification of BB173 162mer adding C-tag QSTMMS

TM3 NST BB173

 

TTA TCA GGA CAT CAT GGT GCT ATT AAG CAT ACT AGG ATC

Amplification of BB173 278mer adding C-tag NST

TM3 QST BB173

 

TTA TCA GGA CAT CAT GGT GCT CTG AAG CAT ACT AGG ATC

Amplification of BB173 278mer adding C-tag QSTMMS

bb0170 qPCR F

 

GTT AAA CCG ATT CCT GGA GAG

Amplification of bb0170 from cDNA for expression studies

bb0170 qPCR R

 

CAG CCA AAA CTT GAT GCT GC

bb0171 qPCR F

 

AAA TCC ATG TCT TTA ATG

Amplification of bb0171 from cDNA for expression studies

bb0171 qPCR R

 

AAC CCT CTC AAG ATT TTC

bb0172 qPCR F

 

TAT GGG GAC AAT TCT TAT ATT CAA

Amplification of bb0172 from cDNA for expression studies

bb0172 qPCR R

 

CAA TCC CAA CCA CAA AAC TTT CCA

bb0173 qPCR F

 

ATT TAA TGA GCC TTT ATA TTT GTT TTT A

Amplification of bb0173 from cDNA for expression studies

bb0173 qPCR R

 

GAT CCA TAA TAT AAA TAT CAT CTA GCT T

bb0174 qPCR F

 

GAT GGT GAA GAG TTT TCC

Amplification of bb0174 from cDNA for expression studies

bb0174 qPCR R

 

TTC TGT TGT AGT GAT TGC

bb0175 qPCR F

 

TTT CAT GAG TTT AGG CCG

Amplification of bb0175 from cDNA for expression studies

bb0175 qPCR R

 

TGT TGA CTT GCT AAA CCC

bb0176 qPCR F

 

TTA CTT GAA GGG GTT CCG

Amplification of bb0176 from cDNA for expression studies

bb0176 qPCR R

 

ATC CCT TTC ACG GAG TGC

rpoN qPCR F

 

TTG TAC CCC ATT CGC ATT TT

Amplification of rpoN from cDNA for expression studies

rpoN qPCR R

 

GTG AAA ACC CCC AAA AAC AA

rpoS qPCR F

 

TTG GGC GAT TTT TCT TCT TC

Amplification of rpoS from cDNA for expression studies

rpoS qPCR R

 

TGC GGG TAA AGG GTT AAA AA

rrp2 qPCR F

 

TGT AGC TTC TCC CCC AAC AC

Amplification of rrp2 from cDNA for expression studies

rrp2 qPCR R

 

TTT TGG CCA TGA AAA AGG AG

bb0420QF

 

TGG CAA GTC GAA TAC CCT CT

Amplification of hpk1 from cDNA for expression studies; From Rogers et al., 2009 [34].

bb0420QR

 

TGT TCG ATT TTA TTG GGA TGC

bb0419F-RT

 

TTG AGG TTG CAA CAA ATG GA

Amplification of rrp1 from cDNA for expression studies; From Rogers et al., 2009 [34].

bb0410R-RT

 

CGG GAT CGC TTT TTA GCT TT