SDS-PAGE of Whole-Cell Proteins and Random Amplified Polymorphic DNA (RAPD) Analyses of Leptotrichia Isolates

Author： Eribe Emenike Ribs K. Eribe Emenike Ribs K.

ISSN： 0891-060X

Source： Microbial Ecology in Health and Disease, Vol.14, Iss.4, 2003-01, pp. : 193-203

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Abstract

In a polyphasic approach to improve the taxonomy of Leptotrichia we have recently demonstrated heterogeneity in the enzymatic/biochemical reactions and cellular fatty acid content of Leptotrichia isolates. In the present study, SDS-PAGE of whole-cell proteins and random amplified polymorphic DNA (RAPD) analyses were used to further characterize and distinguish these isolates of which 57 had been assigned as Leptotrichia buccalis and two as 'Leptotrichia pseudobuccalis'. The phylogenetically closely related Sebaldella termitidis^T and Fusobacterium perfoetens^T were included for comparison. Two major protein bands at approximately 30-40 and 70-80 kDa, respectively, were characteristic for almost all Leptotrichia strains examined as well as for S. termitidis^T while F. perfoetens^T exhibited only the 70-80 kDa major band. Numerical data analysis of the gel patterns obtained with SDS-PAGE assigned most of the strains into 12 clusters at 70% similarity. On RAPD gels with the primer OPA 3 most of the strains shared only one band at approximately 0.40-0.50 kb. With OPA 5 almost all the strains showed two major bands at approximately 0.40 and 0.85 kb, respectively. Numerical analysis of RAPD gels at 70% similarity assigned most strains into ten clusters when the OPA 3 had been used and into 14 clusters with OPA 5. Accordingly, both SDS-PAGE and RAPD demonstrated considerable heterogeneity among Leptotrichia isolates. OPA 5 provided better strain distinction than did OPA 3.