Isolation and characterization of the UDP-glucose 4′-epimerase-encoding gene, galE, from Brucella abortus 2308

• Published in: Gene 1997-11, Vol.202 (1), p.53-59
• Main Authors: Scupham, Alexandra J, Triplett, Eric W.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - isolation & purification; Base Sequence; Benzenesulfonates; Brucella abortus - cytology; Brucella abortus - enzymology; Brucella abortus - genetics; exoB, Sinorhizobium meliloti; Exopolysaccharide synthesis; Fluorescent Dyes; Gene Dosage; Genes, Bacterial; Molecular Sequence Data; Phenotype; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Succinoglycan; UDPglucose 4-Epimerase - chemistry; UDPglucose 4-Epimerase - genetics; UDPglucose 4-Epimerase - isolation & purification
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/S0378-1119(97)00453-8

The UDP-glucose 4′-epimerase-encoding gene, galE or exoB, was isolated from Brucella abortus 2308 by complementation of an exoB mutant of Sinorhizobium meliloti. Confirmation of the identity was done by constructing an in-frame deletion of 660 bp with galE of the B. abortus genome by marker exchange. The resulting galE mutant lacked UDP-glucose 4′-epimerase activity. This activity was restored by in trans complementation with the intact gene. The B. abortus gal E mutant is not altered in colony morphology compared to wt 2308. The lack of UDP-glucose 4′-epimerase activity in the mutant and PCR analysis strongly suggest that only one copy of galE exists in B. abortus 2308. The galE sequence of B. abortus 2308 is more similar to galE from other animal-inhabiting bacteria than it is to exoB from the Sinorhizobium legume symbionts. We propose that galE in B. abortus evolved by lateral transfer from other animal-inhabiting bacteria rather than from a common ancestor of Brucella and Sinorhizobium.