Table 1 Bacterial strains and plasmids.

Bacterial strains

XL1 Blue

recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F´ proAB lacI^qZ∆M15 Tn10 (Tet^R)]

Agilent Technologies

BL21(DE3)

E. coli B F^–ompT gal [E. coli B is naturally dcm and lon] hsdS_B with DE3, a λ prophage carrying the T7 RNA polymerase gene and lacI^Q

Sigma-Aldrich

MG1655

ilvG rpb-50 rph-1 DlacU169

E. coli Genetic Stock Center

WL400

Cam^R Strep^RaraD139 Δ (argF-lac)U169, ptsF25 deoC1 relA1 flbB5301 rpsL150 ΔselD204::cat, conjugation recipient

W. Leinfelder, unpublished data

Plasmids

pLEMO

pACYC184-PrhaBAD-lysY (Cam^R)

⁴⁰

pHT

Kan^R; pET24d derivative T7 expression vector with N-terminal 6xHis-tag and TEV protease cleavage site

¹⁶

pHT TraE

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE

¹⁶

pKM101ΔTraE

Spec^R; pKM101 containing a deletion of the traE locus

Jay E. Gordon & Peter J. Christie, unpublished data

pHT TraE R37A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change R37A

This work

pHT TraE W40A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change W40A

This work

pHT TraE Y106A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change Y106A

This work

pHT TraE R110A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change R110A

This work

pHT TraE Y113A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change Y113A

This work

pHT TraE R176A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change R176A

This work

pHT TraE F177A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change F177A

This work

pHT TraE W195A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change W195A

This work

pHT TraE Y201A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change Y201A

This work

pHT TraE Y203A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change Y203A

This work

pHT TraE R213A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change R213A

This work

pHT TraE F220A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change F220A

This work

pHT TraE Y225A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change Y225A

This work

pHT TraE R226A

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraE with amino change R226A

This work

pHT TraD

Kan^R; T7 promoter vector for the expression of 6xHis-tagged full-length domain of pKM101 TraD

¹⁶

pMSP1D1

Kan^R; pET28a derivate containing mutant (1–11) of MSP1; His-tag on N-terminus followed by spacer sequence and TEV protease site

Addgene

#20,061

pMSP1E3D1

Kan^R; pET28a derivate containing extended MSP1D1; contains repeats of helices 4, 5 and 6; N-terminal 7-his tag followed by spacer sequence and TEV protease cleavage site

Addgene

#20,066

pMSP2N2

Kan^R; pET28a derivate containing tandem MSP; His-tag on N-terminus followed by spacer sequence and TEV protease site

Addgene

#29,520