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Similarity of Ribosomal Proteins Studied by Two-Dimensional Coelectrophoresis for Identification of Gram-Positive Bacteria

Kyowa Hakko Kogyo Co., Ltd., Tokyo Research Laboratories, 3-6-6 Asahimachi, Machidashi, Tokyo, Japan

^* Corresponding author.

ABSTRACT

A two-dimensional electrophoresis method was devised for comparing ribosomal proteins from different bacteria. First-dimension separation of the proteins was carried out by nonequilibrium pH gradient gel electrophoresis in glass capillary tubes. Next, the first-dimension gels were developed in a slab gel by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Within about 3 h (total running time), 40 to 48 distinct and compact ribosomal protein spots were detected in each of the 34 strains belonging to eight genera of gram-positive bacteria which we studied. Similarity coefficients (S_AB values) among the proteins of Corynebacterium and Arthrobacter strains were calculated by determining the numbers of coincident spots obtained by coelectrophoresis. The S_AB values between strains having DNA relatedness values of more than 61% (strains of the same species) were greater than 0.85, whereas the S_AB values between strains of different species were less than 0.45. Ribosomal protein analysis by our improved method is a potential tool for species identification of bacteria.