European Journal of Experimental Biology Libre accès

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Isolation, cloning and transformation studies of Glucan binding protein (GBP) from Streptococcus mutans

Chandrashekhar G. Unakal, V. S. Goudar; A. Naregal,  H.V Pavan, R.M. Yendigery2, Gizachew Yismaw,Mengistu Endris

Many species of bacteria synthesize a polymers and glucan-binding proteins. Less attention has been given to the biological roles of glucans in commensal oral bacteria. The ability of oral streptococci to exploit glucan-binding properties of extracellular and cell-surface proteins to facilitate colonization and survival in the oral environment may have ecological implications for determining the bacterial composition of dental plaque. The transformation studies of gbp gene was done using PUC 18 vector followed by the Sequence analysis. The sequence obtained by PCR (1225 bp) was analyzed by nBLAST at NCBI. The DNA was isolated from the fresh culture, found to have molecular weight more than 24 kb. The sequence analysis confirmed that, the gbp gene as of S. mutans isolate SJ-32 gbp gene. The phylogenetic tree obtained and sequence was further translated to protein to know coding sequences by ORF finder. The virtual protein was further analyzed with pBLAST which confirmed the protein as glucan binding protein of S. mutans. The amplified gene product was transformed into E. coli BL-21 strain. The ability to synthesize extracellular glucans is generally believed to be one of the virulence properties of Streptococcus mutans which contributes to plaque formation and to the subsequent development of dental caries