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Biomedical subjects

R W Davis

Publications and source records attributed to R W Davis.

At least 235 records · Page 13Linked to original sources
PubMed1977

The effects of Escherichia coli and yeast DNA insertions on the growth of lambda bacteriophage.

The effects of Eco RI endonuclease-cleaved Escherichia coli and yeast (Saccharomyces cerevisiae) DNA fragments on the propagation of the lambda bacteriophage vectors containing them were determined on a nonmutanted and a PolA E. coli K12 host. Observable alterations in the growth of hybrids containing yeast DNA insertions were less frequent and less extreme than those seen in hybrids containing E. coli DNA. A lambda-E. coli hybrid was selected after extensive growth on the Pol A (deficient in polymerase I) host which also grew very well on the PolA+ host and may have resulted from some alteration in the hybrid. Hybrids selected on the PolA host gave no evidence for the expression of polymerase I activity. No lambda-yeast hybrid made from the lambdagt vector lacking lambda-specific recombination (red-) had a yield of viable bacteriophage on infection greater than two-thirds that of "wild-type" lambda.

Coliphages↗
PubMed1977

Production of a functional eukaryotic enzyme in Escherichia coli: cloning and expression of the yeast structural gene for imidazole-glycerolphosphate dehydratase (his3).

A cloned segment of yeast DNA containing the structural gene for imidazoleglycerolphosphate dehydratase (D-erythro-imidazoleglycerolphosphate hydro-lase, EC 4.2.1.19) is transcribed and translated in Escherichia coli with sufficient fidelity to produce functional enzyme. This segment of yeast DNA was isolated as a viable molecular hybrid of bacteriophage lambda (lambdagt-Sc2601) which complements a nonrevertible hisB auxotroph of E. coli lacking dehydratase activity. The equivalent segments of DNA cloned from two independent his3 mutants of yeast lacking IGP dehydratase activity do not complement the hisB auxotroph. The two nonfunctional his3 alleles cloned in bacteriophage lambda can be recombined in E. coli to generate a hybrid phage which complements the hisB auxotroph. The dehydratase activity produced in E. coli by the cloned segment of yeast DNA strongly resembles the activity found in yeast.

DNA, Recombinant↗
PubMed1977

Analysis of chromosomal integration and deletions of yeast plasmids.

Plasmid DNAs from six strains of Saccharomyces cerevisiae were compared. Three different plasmids were found, designated Scp 1, Scp 2 and Scp 3, with monomer lengths of 6.19, 6.06 and 5.97 kilobases as referenced to sequenced phiX174 DNA. DNA from each of the plasmids was inserted into a lambda vector DNA. Hybrid phage containing inserted DNA of the desired size were enriched by genetic selection and their DNAs analysed by rapid techniques. All three plasmids share the same organization, two unique sequences separated by two inverted repeats, and share basically the same DNA sequences. Scp 2 and Scp 3 differ from Scp 1 by missing a unique HpaI site and by having small overlapping deletions in the same region. The HpaI site in Scp 1 is, therefore, in a nonessential region and suitable for insertion of foreign DNA in the potential use of the yeast plasmid as a vector. Hybridization of labelled cloned plasmid DNA to restriction fragments of linear yeast DNA separated on agarose gels showed that the plasmid DNA was not stably integrated into the yeast chromosomal DNA.

Chromosomes↗
PubMed1977

Chemical Composition and Ultrastructure of Suberin from Hollow Heart Tissue of Potato Tubers (Solanum tuberosum).

The disorder of potato tubers (Solanum tuberosum var. Russet Burbank) called "hollow heart" is manifested by the occurrence of hollow regions in internal parts of the tuber. The structure and composition of the suberin from the tissue lining of these internal cavities were determined by gas chromatography and mass spectrometry of the LiAlH(4)-hydrogenolysis products. Identification of octadecene-1,18-diol as the major component and the presence of hexadecane-1,16-diol and very long chain (>C(18)) alcohols in the hydrogenolysate showed that the suberin lining the internal cavities is quite similar to that found in the periderm of external wounds and the natural skin. Electron microscopic examination showed similar lamellar structure for the suberin of hollow heart, external wound periderm, and the natural skin of potato tubers. The results show that suberin can develop in a tissue which is not exposed to the external environment.

Journal Article↗
PubMed1977

Comparison of two methods for analysis of aflatoxin M1 in milk.

The cellulose chromatography method of the Association of Official Analytical Chemists for determination of Aflatoxin M1 was compared with a two-dimensional thin layer chromatography procedure. The minimum detectable Aflatoxin M1 by the Official procedure was approximately .01 mug/kg while the thin layer chromatography procedure has a claimed sensitivity of .004 mug/kg. Analyses of milk samples by thin layer chromatography agreed with results of the Official procedure in the range of .01 to .25 mug/liter. Each method consistently yielded recoveries greater than 87% and there was no significant difference between means or variances of each method. The thin layer chromatography method took less time than the Official method and showed a reproducible lower threshold sensitivity of .005 mug/liter.

Aflatoxins↗
PubMed1976

Isolation of bacteriophage lambda containing yeast ribosomal RNA genes: screening by in situ RNA hybridization to plaques.

We have developed an in situ hybridization technique which can be used to screen large numbers of hybrid bacteriophage for the presence of a particular inserted DNA sequence. Plaques of hybrid phage are formed on E. coli lawns on nitrocellulose filters, and their DNA is released, denatured, and fixed directly on the filters for hybridization to radioactive RNA probes. We have used this technique to isolate a number of hybrid bacteriophage lambda which contain EcoRl restriction fragments of the ribosomal RNA genes from yeast, and have examined the DNA from several of these phage.

Coliphages↗
PubMed1976

Functional genetic expression of eukaryotic DNA in Escherichia coli.

We have isolated a segment of DNA from the eukaryote Saccharomyces cerevisiae (baker's yeast) as a viable molecular hybrid of bacteriophage lambda DNA which, when integrated into the chromosome of an E. coli histidine auxotroph, allows this bacterium to grow in the absence of histidine. The nonrevertable, histidine auxotroph lacks the enzymatic activity of imidazole glycerol phosphate (IGP) dehydratase (EC 4.2.1.19). From genetic experiments, we conclude that expression of the segment of yeast DNA results in the production of a diffusible substance and that transcription necessary for the complementation is most likely initiated from the segment of eukaryotic DNA.

Acetoxyacetylaminofluorene↗
PubMed1976

Hybridization of RNA to double-stranded DNA: formation of R-loops.

RNA can hybridize to double-stranded DNA in the presence of 70% formamide by displacing the identical DNA strand. The resulting structure, called an R-loop, is formed in formamide probably because of the greater thermodynamic stability of the RNA-DNA hybrid when it is near the denaturation temperature of duplex DNA. The rate of R-loop formation is maximal at the temperature at which half of the duplex DNA is irreversibly converted to single-stranded DNA (the strand separation temperature of tss) of the duplex DNA and falls precipitously a few degrees above or below that temperature. This maximal rate is similar to the rate of hybridization of RNA to single-stranded DNA under the same conditions. At temperatures above the tss the rate is proportional to the RNA concentration. However, at temperatures below tss the rate of R-loop formation is less dependent upon the RNA concentration. Once formed, the R-loops display considerable stability; the formamide can be removed and the DNA can be cleaved with restriction endonucleases without loss of R-loop structures.

Coliphages↗
PubMed1975

Escalation of aggression: experimental studies.

A finding commonly obtained in research using the Buss "aggression machine" is a main effect for trail blocks, indicating an escalation in shock intensity over trails. Theoretical explanations for this effect were tested in a modified verbal operant-conditioning situation. In Experiment 1, subjects could administer any of 10 levels of positive reinforcement to a "learner" for correct verbal responses or any of 10 levels of negative reinforcement to a learner for incorrect responses. Half of the subjects were required to begin with weak, half with strong, reinforcements. Results indicated that, regardless of condition, subjects gave more intense reinforcements as the learning trails progressed. Those who administered negative reinforcements devalued the learner relative to those who administered positive reinforcements. In Experiment 2, a role-playing procedure was used in which subjects administered either positive or negative reinforcements to a learner whose performance either did or did not improve over trials. Again, in all experimental groups, subjects administered increasingly intense reinforcements over trials. The results are interpreted as supporting a disinhibition theory of anti- and prosocial behavior.

Aggression↗
PubMed1975

T antigen binds to simian virus 40 DNA at the origin of replication.

A technique employing ferritin-conjugated antibody has been developed to visualize specific protein-DNA complexes in the electron microscope and has been used to demonstrate the preferential binding of simian virus 40 (SV40) T antigen at or near the origin of replication of SV40 DNA, 0.67 fractional length clockwise from the EcoRI restriction endonuclease cleavage site. urified covalently closed supercoiled circles of SV40DNA were treated with partially purified T antigen and the complex was stabilized by crosslinking with glutaraldehyde. Hamster antiT antigen gamma-globulin, ferritin-labeled goat anti-hamster gamma-globulin, and glutaraldehyde were then added sequentially. The location of the bound ferritin cores was measured with respect to the EcoRI cleavage site and the orientation of the cores relative to the ends of the DNA was determined with respect to the locations of Escherichia coli DNA unwinding protein, which binds to covalently closed supercoiled SV40 DNA at either of two preferred sites, 0.46 or 0.90 fractional length clockwide from the EcoRI cleavage site.

Antigens, Viral↗
PubMed1975

In vitro construction of bacteriophage lambda carrying segments of the Escherichia coli chromosome: selection of hybrids containing the gene for DNA ligase.

DNA from lambdagt-lambdaB bacteriophage was cleaved with EcoRI endonuclease and fragments from EcoRI-digested E. coli DNA were inserted. This DNA was used to infect E. coli, and phages containing the gene for DNA ligase were isolated by genetic selection. Two different hybrids were found with the same E. coli segment inserted in opposite orientations. Both hybrids produced similar levels of ligase as measured in crude extracts of infected cells.

Chromosome Mapping↗