ABSTRACT
The Escherichia coli K-12 sad gene, which encodes an NAD-dependent succinic semialdehyde dehydrogenase, was cloned into a high-copy-number vector. Minicells carrying a sad+ plasmid produced a 55,000-dalton peptide, the probable sad gene product.
Subject(s)
Escherichia coli/genetics , Genes, Bacterial , Hydroxybutyrate Dehydrogenase/genetics , Bacterial Proteins/analysis , Bacterial Proteins/biosynthesis , Bacterial Proteins/genetics , Chromosome Mapping , Cloning, Molecular , Cosmids , DNA, Bacterial/genetics , Escherichia coli/enzymology , Gene Expression Regulation , Hydroxybutyrate Dehydrogenase/biosynthesis , Mutation , Nucleic Acid Hybridization , Plasmids , gamma-Aminobutyric Acid/analogs & derivatives , gamma-Aminobutyric Acid/metabolismABSTRACT
Escherichia coli strains lacking the terminus region of the chromosome (min 29-36) due to an IS10-promoted deletion did not grow well in rich medium; they also did not grow on fumarate minimal medium because fumAC (min 35.7) is deleted. Strains with secondary mutations that partially suppress the deletion phenotype displayed healthier growth on rich medium and grew on minimal fumarate medium. These suppressor mutants had an IS10 insertion just upstream of the fumB structural gene (min 93.4). A strain with a Tn10 insertion at this location was constructed and used to delete nonessential fumB; fumB deletion mutants grew well on both rich and minimal fumarate media.
Subject(s)
Escherichia coli/genetics , Fumarate Hydratase/genetics , Mutation , Autoradiography , Chromosome Mapping , DNA Transposable Elements , DNA, Bacterial , Genetic Complementation Test , Nucleic Acid Hybridization , Suppression, GeneticABSTRACT
Axon segments isolated from their somata degenerate within days or months depending on species and neuronal type. To better understand the time course of morphological and physiological changes associated with degeneration of axon segments of vertebrate central neurons, we have studied the goldfish Mauthner axon (M-axon) when it has been separated from its soma by spinal cord crush. M-axon segments survive morphologically for at least 77 days at 14 degrees C. Cross-sectional areas of isolated M-axon segments (measured 25-30 mm caudal to the wound site at postoperative days 64 and 77) were greater than those of control axons at the same level. Sheath areas did not change. Electron microscopic observations at the same spinal cord location indicated no clear changes in the configuration or number of neurofilaments or any other organelle. M-axon segments studied morphologically after 87 postoperative days had all degenerated. Mauthner axon segments were capable of conducting action potentials and eliciting ipsilateral EMG responses. Repetitive firing of the M-axon segments elicited EMG responses that fatigued more easily and remained fatigued over a longer interval than did those of control axons. The long duration of M-axon segment survival is unusual in a vertebrate and may be due to the low temperature at which the experiments were conducted (14 degrees C) and/or temperature-independent factors. The increased susceptibility to synaptic depression, which has not reported previously, may represent an early sign of the degenerative process.