TABLE 1.

Characterization of the mutants^a

Functional family	Strain	Gene	Putative function	ORF no.	Mouse CI	Log CFU in HeLa cells/log CFU in macrophages
Transport	19C3	macA	Macrolide efflux	BMEI0359	1.0 × 10−3	2.4 ± 0.1/2.4 ± 0.1
	16H1	tig	Trigger factor	BMEI1069	4.0 × 10−3	2.2 ± 0.1/2.5 ± 0.4
	9A12	ugpA	Glycerol transport	BMEII0591	1.1 × 10−2	1.8 ± 0.2/2.9 ± 1.1
	17F2	ugpA	Glycerol transport	BMEII0624	3.5 × 10−3	2.5 ± 0.1/2.5 ± 0.1
Metabolism	15B2	dppA	Dipeptide uptake	BMEI0433	3.2 × 10−4	2.1 ± 0.4/3.3 ± 0.5
	13D6	ilvC	Val, Leu, and Ile synthesis	BMEI0624	3.1 × 10−2	ND/NDb
	8C7	cobB	Cobalamin synthesis	BMEI0705	4.6 × 10−2	1.8 ± 0.2/3.3 ± 0.8
	8A8	mosA	Inosamine methylase	BMEI1301	3.3 × 10−2	1.7 ± 0.1/2.4 ± 0.6
	16A11	galcD	d-Galactarate dehydratase	BMEII0485	2.2 × 10−3	2.4 ± 0.1/3.0 ± 0.8
	15H1	glcK	Glycerol kinase	BMEII0823	3.3 × 10−2	2.3 ± 0.3/3.1 ± 0.2
Regulation	13B1	asnC family	Transcriptional regulator	BMEI0357	5.4 × 10−4	0.9 ± 0.2/1.8 ± 0.2
	16B12	feuQ	Sensor	BMEI1336	8.0 × 10−4	2.4 ± 0.3/2.4 ± 0.1
	16D3	gntR family	Transcriptional regulator	BMEII1066	1.0 × 10−1	2.4 ± 0.3/2.8 ± 0.1
DNA-RNA metabolism	9A3	rpoA	RNA polymerase α-subunit	BMEI0781	1.2 × 10−4	1.3 ± 0.1/0.5 ± 0.2
	19A4	xseA	Exodeoxyribonuclease	BMEII0527	3.7 × 10−3	1.2 ± 0.2/1.8 ± 0.3
LPS	17F3	pmm	Core biosynthesis	BMEI1396	5.7 × 10−3	2.3 ± 0.3/2.2 ± 0.2
Flagella	9C6	fliF	MS ring	BMEII0151	6.6 × 10−2	2.0 ± 0.3/2.6 ± 0.7
Oxidoreduction	16B2	pheB	Catechol dioxygenase	BMEII0136	3.2 × 10−4	ND/ND
	19A2	fdhA	Formate dehydrogenase	BMEII0378	1.3 × 10−2	ND/ND
	14G4	norE	Nitric oxide reduction	BMEII1001	2.2 × 10−3	1.9 ± 0.1/2.5 ± 0.1
Unknown	10B11	Int. reg.	Intergenic region	BMEI0330-I0331	4.8 × 10−5	1.7 ± 0.1/2.0 ± 0.1
	9A4	Hypothetical protein	Coenzyme A-transferase III	BMEI0898	1.7 × 10−2	2.5 ± 0.6/2.0 ± 0.5
	8B9	Hypothetical protein	Thioesterase	BMEI1167	2.5 × 10−2	2.5 ± 0.5/2.0 ± 0.2
	15F3	Hypothetical protein	Brucella/Agrobacterium orphan gene	BMEI1229	4.5 × 10−1	ND/ND
	14D3	Hypothetical protein	Brucella/Agrobacterium orphan gene	BMEI1339	3.6 × 10−4	1.5 ± 0.1/3.3 ± 0.2
	17E7	Hypothetical protein	α-Proteobacteria orphan gene	BMEI1361	8.5 × 10−4	ND/ND
	13G7	Hypothetical protein	α-Proteobacteria orphan gene	BMEI1433	3.2 × 10−4	1.9 ± 0.1/2.2 ± 0.1
	14B12	Hypothetical protein	EAL domain	BMEI1448	4.5 × 10−3	2.8 ± 0.1/2.5 ± 0.5
	14B5	Hypothetical protein	Tetratricopeptide repeat domain	BMEI1531	3.0 × 10−2	2.9 ± 0.5/0.8 ± 0.2
	17B2	Hypothetical protein	ERFK/YBIS/YCFS/YNHG family	BMEI1809	1.0 × 10−3	1.7 ± 0.2/2.8 ± 0.2
	8B8	Hypothetical protein	Rhizobiaceae orphan gene	BMEI1844	4.7 × 10−3	1.6 ± 0.4/2.0 ± 0.2
	9H5	Hypothetical protein	Brucella/Mesorhizobium orphan gene	BMEI1879	1.8 × 10−2	1.7 ± 0.2/2.0 ± 0.1
	13G9	Hypothetical protein	Beta-lactamase	BMEII0318	2.5 × 10−4	1.4 ± 0.2/2.4 ± 0.2
	13C9	Hypothetical protein	Dipeptidase domain	BMEII0626	7.8 × 10−4	2.8 ± 0.2/2.4 ± 0.5
	8D7	Hypothetical protein	Transmembrane protein	BMEII0935	1.7 × 10−2	1.9 ± 0.2/1.7 ± 0.2
	17B1	Hypothetical protein	Halo-acid hydrolase domain	BMEII1045	1.3 × 10−2	2.6 ± 0.1/3.3 ± 0.4

^aFor each mutant, homology searches were undertaken, using transposon flanking sequences against the genome of B. melitensis. The ORF number for each identified gene is given, and the putative function is based on homologies found by searching the BLAST database for the protein sequence deduced from each ORF. For the ORFs encoding proteins of unknown function, the domains detected by pfam are given. CIs were calculated relative to the wild-type strain B. melitensis 16M. The CI is defined as the output ratio (mutant/wild type) divided by the input ratio (mutant/wild type). Mouse CIs are the averages of CIs determined for two mice. Attenuation in a cellular model at 48 h postinfection is expressed as the difference in the log CFU between the wild type and the mutant. The standard deviations indicated are based on the mutants; the standard deviation for the wild type was ±0.03 for the two cellular models tested. Cellular infections were done individually for each mutant, and the experiments were performed two times in triplicate, with a ratio of 300 bacteria/cell.

^bND, not done.