Cloning, expression, and occurrence of the Brucella Cu-Zn superoxide dismutase.
Recently, the complete amino acid sequence of a protein expressed in Escherichia coli from cloned Brucella abortus DNA was reported. On the basis of amino acid homology, this protein was identified as a copper-zinc superoxide dismutase (Cu-Zn SOD) (B. L. Beck, L. B. Tabatabai, and J. E. Mayfield, Biochemistry 29:372-376, 1990). We demonstrate in this paper that the sequenced protein is the same as the previously studied salt-extractable protein BCSP20. The plasmid-encoded protein expressed from recombinant E. coli is identical to the Brucella-derived BCSP20 in molecular mass, N-terminal amino acid sequence, and cross-reactivity with homologous and heterologous rabbit sera against either the recombinant gene product or the Brucella-derived protein. A survey of the expression of the Cu-Zn SOD protein in Brucella biovars representing all species was done by Western blotting (immunoblotting) using antisera raised against the recombinant E. coli-derived protein. With the exception of B. neotomae and B. suis biovar 2, the Cu-Zn SOD protein was detectable in all Brucella species and biovars tested, including eight biovars of B. abortus.
National Animal Disease Center, U.S. Department of Agriculture, Ames, Iowa 50010.
B J
BJ
Bricker
L B
LB
Tabatabai
B A
BA
Judge
B L
BL
Deyoe
J E
JE
Mayfield
D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology; D002002:Brucella / Q000201:enzymology / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D003300:Copper / Q000378:metabolism*; D003429:Cross Reactions; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D008970:Molecular Weight; D011817:Rabbits; D015183:Restriction Mapping; D013482:Superoxide Dismutase / Q000235:genetics* / Q000276:immunology; D015032:Zinc / Q000378:metabolism*
D000942:Antigens, Bacterial; D003300:Copper; D013482:Superoxide Dismutase; D015032:Zinc