Table 4.

Bacterial strains, plasmids, and primers used in this study.

Strain, plasmid or oligonucleotide primers	Relevant characteristics	Reference
R. leguminosarum bv. trifolii 24.2	Wild type, Rifr, Nxr	[23]
Rt2440	Rt24.2 derivative carrying rosR with one nucleotide deletion (ΔC177)	[23]
Rt5819	Rt24.2 derivative carrying the mini-Tn5 between 363-364 bp position of pssA	[30]
Rt2472	Rt24.2 derivative carrying the mini-Tn5 between 151-152 bp position of rosR	[30]
Rt2441	Rt24.2 with additional rosR upstream region introduced by pM41 integration, Kmr, Nxr	This work
E. coli
DH5α	supE44 ΔlacU169 (φ80 lacZΔ M15) hsdR17 recA1endA1gyrA96 thi-1 relA1	[67]
S17-1	294 derivative RP4-2Tc::Mu-Km::Tn7 chromosomally integrated	[79]
Plasmids
pK19mobGII	mob, lacZα, gusA, Kmr	[80]
pBBR1MCS-2	mob, lacZα, Kmr	[81]
pB31	pUC19 with 1174-bp BamHI fragment containing Rt24.2 rosR	[23]
pM41	pK19mobGII with 586-bp EcoRI-PstI fragment from pB31 containing the rosR upstream region	This work
pRC24	pRK7813 with 1174-bp BamHI fragment containing rosR of Rt24.2	[23]
pBR24	pBBR1MCS-5 with 1174-bp BamHI fragment containing rosR of Rt24.2	[23]
pEX1	pBBR1MCS-2 with 586-bp EcoRI-PstI fragment containing the upstream region and the first 60 codons for RosR	This work
pEX8	pBBR1MCS-2 with 372-bp EcoRI-XbaI fragment containing the -403 bp to -32 bp rosR upstream region	This work
pEX9	pBBR1MCS-2 with 219-bp EcoRI-XbaI fragment containing the -403 bp to -185 bp rosR upstream region	This work
pEX60	pBBR1MCS-2 with 278-bp (-96 bp to +182 bp) EcoRI-PstI fragment containing the first 60 codons for RosR cloned downstream the vector promoter	This work
pBR28	pBBR1MCS-2 with 820-bp (-96 bp to +724 bp) EcoRI-BamHI fragment containing the full-length rosR cloned downstream the vector promoter	This work
pHC60	Vector with gfp and RK2 stabilization fragment, Tcr	[39]
Oligonucleotide primers	Sequence (5'-3')*
pEP1	ATGCAAGAATTCTGCACAGGAAGC	[23]
pEP5	CGGTCAGGAATTCTAAGAACAGGT	[23]
pEP6	TCGAAACAGGAATTCGATTCCTGC	[23]
pRR1	CGCATTCTAGACATGTGATCTGCT	[23]
pEP8	AACGGCTCTAGACTGACACGCCAAA	[23]
pEP9	TCATGCTCTAGACGATGGCCTCAGT	[23]
rosA	GCGGATCCGCGACTTTACCAGATTTA	[23]
rosB	GTCACGCTCTTCGGAATTCAGGGGT	[23]
rosC	AGGGATCCATTCTAAACCTGTCGGCA	[23]
rosD	TCGGATCCTGTCGGCAAAGCATAAGA	[23]
rosG1	GACGATCGAATTCGGCCGTCTCTT	This work
rosD4	TTGCGGATCCGCAGATGCCGGT	This work
rosD5	ACCACGCCTGGGATCCAGGAAAA	This work

* Sequences for EcoRI, BamHI and XbaI restriction sites are underlined.