Methods for studying the genetics, molecular biology, physiology, and pathogenesis of the streptococci

This text is devoted entirely to methods developed in and for studies of members of the bacterial family streptococcaceae. Many of the studies that have been conducted on the streptococcaceae were initiated because of the diseases they cause, or to enhance their utility from an industrial perspective. However, the results of many of these investigations have demonstrated a complexity among some members of the family that warrants an interest in them in their own right, apart from or in addition to any biomedical or industrial considerations. It is therefore hoped and expected that the advanced methods contained in this book should be of interest to those who work with the streptococci and other Gram-positive organisms, to researchers interested in industrial and medical microbiology and to any researcher who seeks to obtain a better understanding of how microorganisms interact with each other, their environment and their hosts.

Table of Contents:
Tn917 transponson mutagenesis and marker rescue of interrupted genes of Streptococcus mutans.- Site-specific homologous recombination mutagenesis in group B streptococci.- Targeted mutagenesis of enterococcal genes.- A lactococcal pWV01-based integration toolbox for bacteria.- Vectors containing streptococcal bacteriophage integrases for site-specific gene insertion.- Streptococcal integration vectors for gene inactivation and cloning.- Induction of transformation in streptococci by synthetic competence stimulating peptides.- Characterization of the lactococcal conjugative element pRS01 using IS946-mediated mutagenesis.- Use of electroportation in genetic analysis of enterococcal virulence.- Genetic transfer methods for Streptococcus sobrinus and other oral streptococci.- Isolation of enterococcal antigen-encoding genes from genomic libraries.- A simple microtiter plate screening assay for bacterial invasion or adherence.- End-probing: A non-radioactive approach to mapping transponson insertions.- A method for mapping phage-inducible promoters for use in bacteriophage-triggered defense systems.- Secretion of heterologous proteins by genetically engineered Streptococcus gordonii.- Examination of streptococcal gene expression in the mammalian environment.- Analysis of adherence-associated gene expression in Streptococcus parasangusis: A method for RNA isolation.- Development of an integrative, lacZ transcriptional-fusion plasmid vector for Streptococcus mutans and its use to isolate expressed genes.- Use of proteomics and PCR to elucidate changes in protein expression in oral streptococci.- The use of continuous flow bioreactors to explore gene expression and physiology of suspended and adherent populations of oral streptococci.- In vitro systems for investigating group Bstreptococcal: host cell and extracellular matrix interactions.- The rat model of endocarditis.- Lipoproteins and other cell-surface associated proteins in streptococci.- Growth of Streptococcal mutons in an iron-limiting medium.- Identification of oral streptococci using PCR-based, reverse-capture, checkerboard hybridization.- Pulsed-field gel electrophoresis as an epidemiologic tool for enterococci and streptococci.- Cell-based panning as a means to isolate phage display Fabs specific for a bacterial surface protein.