A glutamine-amidotransferase-like protein modulates FixT anti-kinase activity in Sinorhizobium meliloti

BMC Microbiology

Table 1 Bacterial strains and plasmids used

Material	Characteristics	References
Strains	Genotype
S. meliloti
GMI211	SmR Nod⁺ Fix⁺	[20]
GMI401	GMI211 asnO::Tn5 SmR NmR	This work
GMI5704	GMI211 fixJ2.3::Tn5 SmR NmR BleoR	[18]
E. coli
ER	asnA31 asnB32 thi	[15]
MM294	Pro-82 thi-1 hsdR17 supE44 endA1	[21]
DH5 α	endA1 hsdR17 (r_k^-mk⁺) supE44 thi-1 recA1 gyrA relA1Δ (lacIZYA-	[29]
	argF)U169 deoR (φ 80dlacΔ (lacZ)M15)
Plasmids
pMF457	pGD926 (IncP broad host range vector) derivative carrying a fixK-
	lacZ fusion with a mutation in the fixT promoter. TcR
pCHK57	pGMI41211 (Inc-P1 broad host range vector) derivative carrying a	[31]
	nifA-lacZ fusion. TcR
pMF11	Same as pMF10 but with fixT in reverse orientation. GmR
pBH1	pBBR1-MCS3 derivative, carrying an HindIII fragment containing	This work
	the asnO gene and the SMb20482 orf.
pBasn2	pBBR1-MCS3 derivative, expressing asnO under the control of the	This work
	pLac promoter
pUC23	pUC18 digested with EcoRI, carrying the S. meliloti EcoRI	This work
	fragment containing the asn O region with the Tn5 insert. Ap^R,
	Km^R.
BAC37	pBeloBAC11 containing a 100 kb S. meliloti genome fragment	[26]
	carrying the asn-like region of pSymB.
pRK602	PRK600::Tn5, Cm^r, Nm-Km^r	[21]