DNA sequence and in vitro mutagenesis of the gene encoding the fructose-1,6-diphosphate-dependent L-(+)-lactate dehydrogenase of Streptococcus mutans.
Previously, the fructose-1,6-diphosphate-dependent L-(+)-lactate dehydrogenase gene of Streptococcus mutans JH1000 was cloned into Escherichia coli (J. D. Hillman, M. J. Duncan, and K. P. Stashenko, Infect. Immun. 58:1290-1295, 1990). In the present study, the nucleotide sequence of 1.29 kb of S. mutans DNA which contained the promoter and protein-coding region of the gene was determined. In vitro disruption of the gene was achieved by deletion of the promoter and a major portion of the protein-coding sequence. Subsequently, a tetracycline resistance gene from S. mutans was inserted at the deletion site as a marker for selection. In addition, evidence from Southern hybridization showed that S. mutans JH1000 contained a single copy of the lactate dehydrogenase gene.
Department of Molecular Genetics, Forsyth Dental Center, Boston, Massachusetts 02115.
M J
MJ
Duncan
J D
JD
Hillman
D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000096:biosynthesis; D001483:Base Sequence; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000235:genetics*; D005635:Fructosediphosphates / Q000378:metabolism; D005798:Genes, Bacterial; D007770:L-Lactate Dehydrogenase / Q000235:genetics*; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D016415:Sequence Alignment; D013295:Streptococcus mutans / Q000235:genetics*
D001430:Bacteriocins; D004269:DNA, Bacterial; D005635:Fructosediphosphates; D007770:L-Lactate Dehydrogenase; C029063:fructose-1,6-diphosphate