Abstract
The previously cloned gene for L-(+)-lactate dehydrogenase (LDH) from Streptococcus mutans was mutagenized in vitro. An Escherichia coli transformant which expressed a thermolabile LDH activity was identified. The ldh(Ts) gene was introduced into S. mutans on a suicide vector to create a heterodiploid expressing both wild-type and thermolabile LDH activities. Self-recombinants which had only one ldh gene were isolated. One of these clones expressed only the thermolabile LDH activity. This isolate grew well at 30 degrees C but did not grow at 42 degrees C under a variety of cultivation conditions, thereby proving that LDH deficiency is lethal in S. mutans in the absence of compensatory mutations.
MeSH Terms
Blotting, Southern
Chromosomes, Bacterial
Cloning, Molecular
DNA, Bacterial/analysis
Enzyme Stability
Genes, Bacterial
Hot Temperature
Hydroxylamine
Hydroxylamines/pharmacology
L-Lactate Dehydrogenase/chemistry,genetics,metabolism
Mutagenesis
Plasmids/drug effects
Recombination, Genetic
Restriction Mapping
Streptococcus mutans/enzymology,genetics,growth & development
Thermodynamics
Chemicals
DNA, Bacterial
Hydroxylamines
Hydroxylamine
L-Lactate Dehydrogenase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Chen A
Department of Oral Biology, University of Florida College of Dentistry, Gainesville 32610.
Hillman J D
Duncan M
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