Genetic cloning of a protease and complete nucleotide sequence of a neuraminidase from streptococcus oralis C104
Almeida, Anna Galganny
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Streptococcus sanguis, Streptococcus oralis and Streptococcus mitis are among the earliest colonizers of the tooth surface during plaque formation. In addition, these species are also associated with extraoral infections including infective endocarditis. A wide range of proteolytic enzymes produced by S. oralis is believed to play an essential role in plaque formation and systemic diseases. Furthermore, neuraminidases, enzymes that cleave sialic acids from carbohydrates and glycoproteins, are believed to play a role in plaque formation and extraoral infections by oral streptococci. In order to identify the gene(s) encoding the S. oralis neuraminidase, a genomic library from S. oralis C104 was constructed in Lambda ZAP II and screened with MUN (a fluorescent neuraminidase substrate, 2'-(4-methylumbelliferyl)-aD- N-acetylneuraminic acid) to detect plaques with neuraminidase activity. Approximately 500,000 plaques were screened and 15 plaques with neuraminidase activity were obtained. E. coli containing recombinant plasmid obtained by in mvo excision from 9 neuraminidase-positive plaques were subcultured and expressed neuraminidase activity. [TRUNCATED]
PLEASE NOTE: This work is protected by copyright. Downloading is restricted to the BU community: please click Download and log in with a valid BU account to access. If you are the author of this work and would like to make it publicly available, please contact firstname.lastname@example.org.Thesis (D.Sc.D.)--Boston University, Henry M. Goldman School of Dental Medicine, 1999 (Pediatric Dentistry).Includes bibliographical references (p. 81-109).
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