ABSTRACT
The mdx mouse mutation arises from a C-to-T point mutation, which terminates the translation of dystrophin and results in the loss of a functional dystrophin protein. mdx mice are used widely in studies of the role of dystrophin and of potential treatments for Duchenne muscular dystrophy, thus accurate genotyping is essential. Current methods require labor-intensive efforts and can often lead to misconstrued results. This study describes a simple and highly reliable, sensitive, and user-friendly, high-resolution melt (HRM) assay that is able to utilize DNA obtained from a variety of sources in order to genotype the known sequence variant of the mdx mouse. Muscle Nerve 39: 603-608, 2009.
Subject(s)
Dystrophin/genetics , Muscular Dystrophy, Animal/genetics , Point Mutation/genetics , Polymerase Chain Reaction/methods , Animals , Disease Models, Animal , Genotype , Mice , Mice, Inbred C57BL , Mice, Inbred mdxABSTRACT
Seventy-two lactic acid producing bacterial isolates (excluding streptococci) were cultured from the gastrointestinal tract of six horses. Two of the horses were orally dosed with raftilose to induce lactic acidosis and laminitis while the remaining four were maintained on a roughage diet. Near complete 16S rDNA was amplified by PCR from the genomic DNA of each isolate. Following RFLP analysis with the restriction enzymes MboI, HhaI and HinfI, the PCR products from the 18 isolates that produced L- and/or D-lactate were subsequently cloned and sequenced. DNA sequence analysis indicated that the majority of the isolates were closely related to species within the genus Lactobacillus, including Lactobacillus salivarius, Lactobacillus mucosae and Lactobacillus delbrueckii. Four isolates were closely related to Mitsuokella jalaludinii. Lactic acid producing bacteria (LAB) from the equine gastrointestinal tract was dominated by representatives from the genus Lactobacillus, but also included D-lactate-producing bacteria closely related to M. jalaludinii. Identification and characterization of LAB from the equine gastrointestinal tract should contribute to our understanding and management of fermentative acidosis, ulceration of the stomach and laminitis.
