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Table 2 Bacterial strains and plasmids used in this study

From: Role of the small RNA RyhB in the Fur regulon in mediating the capsular polysaccharide biosynthesis and iron acquisition systems in Klebsiella pneumoniae

Strains or plasmids

Descriptions

Reference or source

K. pneumoniae

  

CG43S3

CG43 Smr

[56]

ΔlacZ

CG43S3ΔlacZ

[17]

Δfur

CG43S3Δfur

[22]

ΔlacZ Δfur

CG43S3ΔlacZ Δfur

[22]

ΔryhB

CG43S3ΔryhB

This study

Δfur ΔryhB

CG43S3Δfur ΔryhB

This study

ΔlacZ Δfur ΔryhB

CG43S3ΔlacZ Δfur ΔryhB

This study

ΔgalU

CG43S3ΔgalU

[57]

E. coli

  

DH5α

supE44 ΔlacU169 (f80 lacZ Δμ15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1

[58]

BL21-RIL

F-ompT hsdS B [r B -m B -]gal dcm [DE3]

Laboratory stock

S17-1 λ pir H1717

hsdR recA pro RP4-2 [Tc::Mu; Km::Tn7] [λpir] araD139 ΔlacU169 rpsL150 relA1 flbB5301 deoC1 ptsF25 rbsR aroB fhuF::λ placMu

[59, 60]

Plasmids

  

pKAS46

Positive selection suicide vector, rpsL Apr Kmr

[59]

yT&A

TA cloning vector

Yeastern

pRK415

Broad-host-range IncP cloning vector, Tcr

[61]

pT7-7

Cloning vector, Apr

[62]

pETQ

Kmr, protein expression vector

[61]

placZ15

Cmr, promoter selection vector, lacZ+

[17]

pfur

Tcr, 0.8-kb fragment containing a fur allele cloned into pRK415

[22]

pET30c-Fur

Kmr, 450-bp fragment encoding full-length Fur cloned into pET30c

[22]

pRyhB04

2.0 kb fragment containing an internal ~70-bp deletion in ryhB cloned into pKAS46

This study

pRyhB15

Cmr, 178-bp fragment containing the region upstream of ryhB cloned into placZ15

This study

pOrf12

Cmr, 500-bp fragment containing the region upstream of Klebsiella K2 cps orf1-orf2 cloned into placZ15

[17]

pOrf315

Cmr, 900-bp fragment containing the region upstream of Klebsiella K2 cps orf3-orf15 cloned into placZ15

[17]

pOrf1617

Cmr, 300-bp fragment containing the region upstream of Klebsiella K2 cps orf16-orf17 cloned into placZ15

[17]

pT7-7-pryhB

178-bp fragment containing the putative ryhB promoter, cloned into pT7-7

This study

pETQ-ryhB

Kmr, 326-bp fragment containing the promoter and coding region of ryhB cloned into pETQ

This study