Promotor mutants of the yeast his3 gene.
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Biomedical subjects
Publications and source records attributed to R W Davis.
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Natural dives are usually short and aerobic, involving swimming effort that is not energetically costly. Present evidence indicates few, if any, major biochemical enzyme adaptations that would promote an extended anaerobic capacity in marine mammals. We have discussed the lack of evidence for unusual biochemical adaptations for anaerobiosis and the importance and characteristics of natural aerobic dives. What explanation can we offer for the known capability of some marine mammals, such as Weddell seals and sperm whales, to remain submerged for over an hour? Attributes that serve them well for shorter dives are important also for long dives, such as: (a) a large total oxygen store relative to body size, and (b) parsimonious use of blood oxygen due to the lowered energy requirements of various organs. During long dives, some organs deplete their oxygen stores, become predominately dependent on anaerobic metabolism, and build up large quantities of lactate. The result is a degree of fatigue that leaves the animal incapable of further intense diving activities for some time. Consequently, long anaerobic dives are quite rare in nature.
Cardiac output during cardiopulmonary resuscitation (CPR) was measured by a modified indicator-dilution technique in 20 anesthetized dogs (6-12 kg), during repeated 1- to 2-min episodes of electrically induced ventricular fibrillation, by a mechanical chest compressor and ventilator. With compression rates from 20 to 140/min and compression durations (duty cycles) from 10 to 90% of cycle time, cardiac output (CO) was predicted by the equation: CO = CR . SVmax . [DC/(k1 . CR + DC)] . [(1 -- DC)/k2 . CR + 1 - DC)], where CR is compression rate, DC is duty cycle, SVmax (19 ml) is the effective capacity of the pumping chamber, and k1 (0.00207 min) and k2 (0.00707 min) are ejection and filling constants. This expression predicts maximal CO for DC = 0.40 and cR = 126/min and 90-100% of maximal CO for 0.3 less than DC less than 0.5 and 70 less than CR less than 150/min. Such mathematical analysis may prove useful in the optimization of CPR.
Plasma glucose, glucagon, and insulin concentrations were measured in the harbor seal, Phoca vitulina, during a 6-min dive and a 30-min recovery period. Studies were performed in the fasting state and following intravenous glucose. During diving in the fasting state, values of plasma glucose, glucagon, and insulin are not significantly different from prediving values, presumably because of the loss of perfusion to the pancreas. In the postdiving period, plasma glucagon increases significantly. The increased glucagon levels reach a peak at 6 min into the postdiving period. By 4 min postdiving, plasma glucose values increase. The hyperglycemia appears to be at least partially related to hyperglucagonemia. Despite hyperglycemia, insulin levels do not change significantly. The net effect is to increase glucose availability, particularly for non-insulin-requiring tissues (brain). Preadministration of glucose eliminates the postdiving increase in glucagon. Plasma glucose and insulin concentrations also show no significant changes during the diving and postdiving periods. These results suggest that hormonally mediated glucose conservation serves to maintain brain glucose supplies and to restore peripheral carbohydrate stores during the postdiving period.
Yeast transformation can be used to insert new sequence arrangements into a variety of chromosomal locations by homologous recombination. These newly inserted sequences can recombine with similar sequences located on other chromosomes. In these events, information is duplicated without being lost at the site from which it is derived. Similar mechanisms might be utilized by cells to provide new functions during development or differentiation.
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A selective scheme is presented for isolating sequences capable of replicating autonomously in the yeast Saccharomyces cerevisiae. YIp5, a vector that contains the yeast gene ura3, does not transform a ura3 deletion mutant to Ura+. Hybrid YIp5-Escherichia coli DNA molecules also fail to produce transformants. However, collections of molecular hybrids between YIp5 and DNA from any of six eukaryotes tested (S. cerevisiae, Neurospora crassa, Dictyostelium discoideum Ceanhorabditis elegans, Drosophila melanogaster, and Zea mays) do transform the deletion mutant. The Ura+ transformants grow slowly, are unstable under nonselective conditions, and carry the transforming DNA as autonomously replicating, supercoiled circular molecules. Such a phenotype is qualitatively identical to that of strains transformed by molecules containing a yeast chromosomal origin of replication. Thus, these DNA hybrid molecules may contain eukaryotic origins of replication. The isolated sequences may be useful in determiing the signals controlling DNA replication in yeast and in studying both DNA replication and transformation in other eukaryotic organisms.
We describe a new basis for the construction of a genetic linkage map of the human genome. The basic principle of the mapping scheme is to develop, by recombinant DNA techniques, random single-copy DNA probes capable of detecting DNA sequence polymorphisms, when hybridized to restriction digests of an individual's DNA. Each of these probes will define a locus. Loci can be expanded or contracted to include more or less polymorphism by further application of recombinant DNA technology. Suitably polymorphic loci can be tested for linkage relationships in human pedigrees by established methods; and loci can be arranged into linkage groups to form a true genetic map of "DNA marker loci." Pedigrees in which inherited traits are known to be segregating can then be analyzed, making possible the mapping of the gene(s) responsible for the trait with respect to the DNA marker loci, without requiring direct access to a specified gene's DNA. For inherited diseases mapped in this way, linked DNA marker loci can be used predictively for genetic counseling.
A yeast DNA sequence that behaves as a chromosomal replicator, ars1 (autonomously replicating sequence), has been isolated. On transformation, ars1 allows autonomous replication of all co-linear DNA. The replicator can integrate into other replication units and can function in multimeric form. The 850-base pair ars1 element has no detectable homology to other yeast sequences. Such replicator-containing plasmids can be used for the isolation of DNA sequences in yeast cells as well as for the study of chromosomal DNA replication.
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