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The authors would like to make the following corrections to the above paper.
1. In Table 1
, page 456, strains/plasmids HASF300, pKS : hasF : Gmr, pUCGM, pKNGhasFGmr are added and the characteristics of pEXSH, pEXH, pEXS, pEXR, pKS : sdeB, and pKS : HasF are amended. The correct version of the table is given below.
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(a) Line 40, beginning with, ‘The streptomycin-resistant strains...’, replace with:
The streptomycin-resistant strains (with pKNG101, pKNGsdeB, pKNGhasF, pKNGsdeR, pKNGhasFGmr) were grown on LB plates and in LB broth containing 50 µg streptomycin ml–1 (Sigma-Aldrich).
Continues with ‘The carbenicillin-resistant strains...’
(b) Line 45, additional text:
The gentamicin-resistant strains (with pUCGM, pKS : hasF : Gmr) were grown on LB plates and in LB broth containing 20 µg gentamicin ml–1 (Sigma-Aldrich).
Continues with ‘Strains SDEAB1, SDEAB2, HASF100...’
(c) Line 45, beginning with ‘Strains SDEAB1, SDEAB2, HASF100...’, replace with:
SDEAB1, SDEAB2, HASF100, HASF200 and SDER1 strains were grown on LB plates and in LB broth containing 100 µg ampicillin ml–1 and 50 µg streptomycin ml–1, HASF300 was grown on media containing 20 µg gentamicin ml–1 and 50 µg streptomycin ml–1, and SDEAB3/HASF300 was grown on media containing 25 µg kanamycin ml–1 and 20 µg gentamicin ml–1.
Continues with ‘UOC-67, MT616, CC118...’
3. Construction of S. marcescens hasF-, sdeB- and sdeR-deficient strains, page 455, second column
(a) Line 6, additional text:
For creation of a hasF insertion mutant, a 900 bp PstI-flanked Gmr cassette from pUCGM was inserted into a unique PstI restriction site within the EcoRI/EcoRI flanked hasF fragment of pKS : HasF resulting in pKS : hasF : Gmr. pKS : hasF : Gmr was digested with EcoRI to remove the 2.4 kb disrupted hasF and the ends of the fragment were made flush with T4 DNA polymerase. The disrupted hasF gene was then ligated into the pKNG101 replacement vector resulting in pKNGhasFGmr, which was then transformed into E. coli CC118 in similar fashion as above.
Continues with ‘The sdeB gene (Kumar & Worobec, 2005a) was recloned as...’
(b) Line 6, beginning with ‘The sdeB gene (Kumar & Worobec, 2005a) was recloned...’, replace with:
The sdeB gene (Kumar & Worobec 2005a) was cloned as a 1.7 kb EcoRI/EcoRI fragment into pKS, and transformed into UOC-67 in order to be disrupted by the insertion of a kanamycin resistance cassette into a unique PstI restriction site.
Continues with ‘The sdeR gene (GenBank accession...’
(c) Line 15 of text, beginning with ‘Conjugation between the E. coli...’, replace with:
Conjugation between the E. coli strain harbouring each of the disrupted S. marcescens sdeB, hasF (both insertional and deletional mutations) and sdeR genes and the wild-type S. marcescens strain was carried out using E. coli helper strain MT616 (Finan et al., 1986) to create SDEAB1, HASF100 (deletional mutation), HASF300 (insertional mutation) and SDER1, respectively.
Continues with ‘Conjugation was also carried out...’
(d) Line 23, beginning with ‘For the double sdeB/hasF-deficient...’, replace with:
For the double sdeB/hasF-deficient mutant (SDEAB3/HASF300), conjugation was carried out between E. coli CC118(pKNGsdeB) and S. marcescens (HASF300), the hasF-deficient strain, using E. coli MT616 as a helper.
Continues with, ‘Analysis of transconjugants was based on sucrose...’
4. Complementation of sdeB-, hasF- and sdeR-deficient strains, page 455, second column
(a) Line 34, beginning with ‘For complementation purposes...’, replace with:
For complementation purposes, the expression vector pEX1.8 was used for cloning a 1.5 kb wild-type EcoRI/HindIII hasF fragment (pEXH), a 3.1 kb wild-type EcoRI/HindIII sdeB fragment (pEXS) and a 0.4 kb wild-type EcoRI/HindIII sdeR fragment (pEXR).
Continues with ‘In addition, hasF and sdeB were introduced...’
(b) Line 39, beginning with ‘To complement both hasF and sdeB...’, replace with:
To complement both hasF and sdeB simultaneously, a 3.1 kb EcoRI/EcoRI sdeB fragment was cloned into the EcoRI/EcoRI site, and a 1.5 kb hasF HindIII/HindIII fragment was cloned into the HindIII/HindIII site of pEX1.8 (pEXSH).
Continues with ‘To construct strain SM2000, pEXH was...’
REFERENCES
Finan, T. M., Kunkel, B., De Vos, G. F. & Signer, E. R. (1986). Second symbiotic megaplasmid in Rhizobium meliloti carrying exopolysaccharide and thiamine synthesis genes. J Bacteriol 167, 66–72.
Kaniga, K., Delor, I. & Cornelis, G. R. (1991). A wide-host-range suicide vector for improving reverse genetics in Gram-negative bacteria: inactivation of the blaA gene of Yersinia enterocolitica. Gene 109, 137–141.[CrossRef][Medline]
Kumar, A. & Worobec, E. A. (2005a). Cloning, sequencing and characterization of the SdeAB multidrug efflux pump of Serratia marcescens. Antimicrob Agents Chemother 49, 1495–1501.
Pearson, J. P., Pesci, E. C. & Iglewski, B. H. (1997). Roles of Pseudomonas aeruginosa las and rhl quorum-sensing systems in control of elastase and rhamnolipid biosynthesis genes. J Bacteriol 179, 5756–5757.
Schweizer, H. D. (1993). Small broad-host-range gentamycin resistance gene cassettes for site-specific insertion and deletion mutagenesis. Biotechniques 15, 831–834.[Medline]
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