Identification of several rod loci and cloning of the rodD locus of Streptococcus mutans.
Previous work has shown that Streptococcus mutans is normally a short rod or a sphere, depending on its environment. This paper describes two distinct genetic approaches used to identify multiple loci and isolate one locus, rodD, controlling S. mutans rod shape. The first method involved isolation of a group of rod- mutants caused by transposon Tn916 insertion, and analysis of the inactivated genes by Southern hybridization. The second method involved mutagenesis via a shotgun insertion-duplication technique, isolation of a rod- mutant, and cloning the intact rod locus, employing an integration shuttle plasmid, pVA891. These approaches have led to the identification of multiple rod loci involved in determining the rod shape of S. mutans, and also cloning of one rod locus, rodD. The cloning strategy may also be useful for cloning other streptococcal genes which cannot be detected by their expression in Escherichia coli.
Department of Oral Diagnosis, University of Connecticut, Health Center, Farmington 06030.
L
L
Tao
J M
JM
Tanzer
H K
HK
Kuramitsu
A
A
Das
D015139:Blotting, Southern; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004926:Escherichia coli; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008040:Genetic Linkage; D016254:Mutagenesis, Insertional; D010641:Phenotype; D013295:Streptococcus mutans / Q000166:cytology / Q000235:genetics*
D004251:DNA Transposable Elements