Regulation of surface synthesis in Streptococcus mutans GS-5 will be examined in three systems. In the first system we will look at the regulation of the expression of two cloned genes, gtfB and gtfC. We will use for this purpose a series of apparent deletion mutants that express varying amounts of antigen-reactive and enzymatically active glucosyltransferase. The mutants will be characterized biochemically and by restriction mapping in an effort to pinpoint various regulatory regions. The putative promoter of gtfB will be cloned in a promoter vector and examined for possible effects of sucrose on expression of these gtf genes in S. mutans. We will study serotype c polysaccharide biosynthesis using 4 mutants that were obtained using MAbs generated against different epitopes. We will complement the genetic defects with DNA cloned into a streptococcal vector. In a third series of experiments, we will identify, and characterize surface-associated and supernate proteins produced by S. mutans GS-5 under various growth conditions and will identify and/or generate monoclonal antibodies (MAbs) to these proteins. The MAbs will then be used to isolate and identify Tn9l7-inactivated mutants of S. mutans for various surface proteins. The use of Tn9l7 inactivation will permit mapping of the mutation, rapid cloning of a portion of the inactivated gene, and eventually, molecular characterization.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
2R01DE005180-10A3
Application #
3219268
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Project Start
1979-01-01
Project End
1996-02-29
Budget Start
1991-03-01
Budget End
1992-02-29
Support Year
10
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Temple University
Department
Type
Schools of Medicine
DUNS #
City
Philadelphia
State
PA
Country
United States
Zip Code
19122
Tudor, J J; Marri, L; Piggot, P J et al. (1990) Size of the Streptococcus mutans GS-5 chromosome as determined by pulsed-field gel electrophoresis. Infect Immun 58:838-40
Zito, E T; Daneo-Moore, L (1988) Transformation of Streptococcus sanguis to intrinsic penicillin resistance. J Gen Microbiol 134:1237-49
Procino, J K; Marri, L; Shockman, G D et al. (1988) Tn916 insertional inactivation of multiple genes on the chromosome of Streptococcus mutans GS-5. Infect Immun 56:2866-70
Daneo-Moore, L; Volpe, A (1985) Recombination-deficient Streptococcus sanguis. Infect Immun 48:584-6