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Biomedical subjects

D J Brenner

Publications and source records attributed to D J Brenner.

At least 325 records · Page 18Linked to original sources

Conservation of transfer ribonucleic acid and 5S ribonucleic acid cistrons in Enterobacteriaceae.

The genes for tranfer ribonucleic acid (tDNA) and 5S ribonucleic acid (5SDNA) were isolated from the total deoxyribonucleic acid (DNA) of Escherichia coli. The relatedness of tDNA and 5S from E. coli and other species of Enterobacteriaceae was determined by reassociation of the isolated genes labeled with 32PO4 to unlabeled, unfractionated DNA. Double-stranded DNA was separated from unreacted DNA by hydroxyapatite chromatography. Thermal elution profiles were done to determine the amount of unpaired bases present in related DNA sequences. Relative to total DNA, both 5S DNA and tDNA were highly conserved throughout the Enterobacteriaceae, including the genera Yersinia and Proteus.

DNA, Bacterial↗

Unusual Enterobacteriaceae. "Proteus rettgeri" that "change" into Providencia stuartii.

A blood culture bottle from a patient with bacteremia contained both Proteus rettgeri biogroup 5 and Providencia stuartii (described in Bergey's Manual of Determinative Bactiology [8th ed., 1974] as Proteus inconstans), which had the same unusual antibiotic resistance pattern. Single colonies of this P. rettgeri biogroup 5 isolate were shown to produce urea- clones. If current taxonomy is used, the strain changed from P. rettgeri to P. stuartii in the laboratory and probably also in the patient. Urea- clones were also found in three of six other strains of P. rettgeri biogroup 5. No urea-negative clones were found in two isolates each of P. rettgeri biogroups 1 and 3. Previous data from deoxyribonucleic acid-deoxyribonucleic acid hybridization, biochemical reactions, and serological cross-reactions all have indicated that the taxon now called P. rettgeri biogroup 5 should be classified as P. stuartii urea+. We propose that this taxonomic change be made. Urease production is probably plasmid mediated in P. stuartii urea+ and can easily be lost, as shown in our case report and in three stock cultures. Urea hydrolysis will no longer be the key test for differentiating P. rettgeri from P. stuartii. Rather, acid production from trehalose, D-arabitol, adonitol, and D-mannitol will be the key tests. Whereas P. rettgeri is usually trehalose-, D-arabitol+, adonitol+, and D-mannitol+, P. stuartii has the opposite reactions.

Aged↗

DNA relatedness among species of Enterobacter and Serratia.

Species of Enterobacter and Serratia were examined for deoxyribonucleic acid relatedness to Klebsielleae, to atypical erwiniae, and to other members of Enterobacteriaceae. Deoxyribonucleic acid hybridization and then hydroxyapatite chromatography was the technique used to assess relatedness. Strains of Enterobacter cloacae formed two separate hybridization groups that correlate with the presence or absence of yellow pigment. Pigmented E. cloacae were 75-100% related, but they were only 40-50% related to unpigmented strains. Conversely, unpigmented strains were 70% or more related but were only 40-50% related to the pigmented strains. Both pigmented and unpigmented E. cloacae were 40-45% related to Enterobacter aerogenes and klebsiellae, and 20-30% related to Serratia species and Enterobacter hafniae. Atypical erwiniae were highly related to E. cloacae. Serratia marcescens strains formed one closely related group. Serratia liquefaciens strains formed a single, more disperse, relatedness group, as did isolates of Serratia rubidaea. These species were related throughout a substantial portion of their genomes. A group of lysine-positive "Citrobacter-like" strains were 40-50% related to Serratia species. Only four E. hafniae strains were tested. Two of these were highly related, while the other two were only 50% related to the reference strain. Enterobacter hafniae was only 15-20% related to other Enterobacteriaceae.

DNA, Bacterial↗

Shigella flexneri 6 biotypes: a review.

Except in gas production, S. flexneri 6 is generally similar to other S. flexneri. S. flexneri 6 contains both aerogenic and anaerogenic strains which are identical serologically. It is not surprising that S. flexneri 6 is not separable from other S. flexneri types on the basis of DNA relatedness (overall genetic makeup) (4). The Manchester biotype is seen in the United States, and is easily separable from other Enterobacteriaceae by biochemical tests. One can no longer assume à priori that any given biochemical reaction rules an organism out of Shigella. This is particularly true for the production of gas from glucose, but also applies to the fermentation of lactose and the production of H2S. Clinical laboratory personnel should be alert to these possibilities.

Aerobiosis↗

Homology between the deoxyribonucleic acid of fertility factor P and Vibrio cholerae chromosomal deoxyribonucleic acid.

The deoxyribonucleic acid (DNA) of the Vibrio cholerae fertility factor P was isolated by the dye-buoyant density method and hybridized to V. cholerae chromosomal DNA. The DNA of this fertility plasmid had between 35 to 40% homology with the V. cholerae chromosomal DNA. Little or no homology was detected between the P factor DNA and DNA of the Escherichia coli sex factor F.

Centrifugation, Density Gradient↗

Evaluation of the Enteric Analyzer, an instrument to aid in the identification of Enterobacteriaceae.

This study evaluated the Enteric Analyzer, an instrument designed to identify Enterobacteriaceae, with data obtained from the Modified r/b Enteric Differential System, other rapid identification systems, or conventional identification systems. It is programmed for 19 reactions with data obtained from Center for Disease Control publications. The instrument is very simple to use. Typical strains from 25 of 28 species were unequivocally identified. With the other three species, two choices were given. A switch allows the user to identify all species possibilities where a given biochemical pattern occurs in more than 1% of the strains or more than 10% of the strains. The instrument is useful both in the clinical laboratory and for teaching purposes.

Classification↗

Molecular relationships among the Salmonelleae.

Polynucleotide sequence relatedness studies were carried out to determine the extent of divergence present in members of the tribe Salmonelleae and between salmonellae and other enteric bacteria. Typical Salmonella were 85 to 100% related. Two groups of biochemically atypical Salmonella showed somewhat lower binding to typical salmonellae and to each other. Arizona were 70 to 80% related to salmonellae. Two groups of Arizona were detected. These groups correlated with the presence of monophasic or diphasic flagellar antigens. Salmonella and Arizona were no more related to Citrobacter than to Escherichia coli (45-55%). Relatedness of Salmonella and Arizona to other enterobacteria ranged from 20 to 40% with klebsiellae and shigellae, to 20 to 25% with erwiniae, and to less than 20% with edwardsiellae and Proteus mirabilis.

Base Sequence↗

Use of a single-strand specific nuclease for analysis of bacterial and plasmid deoxyribonucleic acid homo- and heteroduplexes.

Bacterial and plasmid homo- and heteroduplexes have been analyzed with a single-strand specific endonuclease, S1, of Aspergillus oryzae. Under appropriate assay conditions, there was a high degree of correlation between the degree of deoxyribonucleic acid (DNA)-DNA homoduplex formation assessed by the S1 endonuclease and by hydroxyapatite (HA). Heteroduplexes which contain extensive regions of polynucleotide sequences in common are similarly recognized by the S1 endonuclease and HA. In instances where there is little or imperfect complementarity between heterologous DNA strands, the S1 endonuclease and the HA method give slightly different estimates. From DNA duplex thermal stability experiments assayed with the S1 endonuclease, there is preliminary evidence that well-matched sequences identified by the enzyme are not similarly recognized by HA. The assay of homo- and heteroduplexes with the S1 endonuclease permits an accurate, reproducible and rapid determination of polynucleotide sequence relationships and may be seriously considered as a method of choice for survey work and for investigations which require a large number of DNA-DNA hybridization assays.

Aspergillus↗

Polynucleotide sequence relatedness among three groups of pathogenic Escherichia coli strains.

Escherichia coli strains that cause dysentery-like disease, parenteral infection, and infantile diarrhea form specific groups based on mobility of O and K antigens in immunoelectrophoresis. Members from each of these groups were assayed for gross nucleotide sequence relatedness. The method used was interspecific deoxyribonucleic acid (DNA) reassociation reactions carried out free in solution. Reassociated DNA was separated from unreacted DNA by passage through hydroxyapatite. DNA relatedness between these groups was approximately 80%. The groups containing those strains causing parenteral infection and those responsible for dysentery-like disease showed preferentially high intragroup DNA relatedness. The group containing strains responsible for infantile diarrhea did not show preferentially high intragroup DNA relatedness with the reference strain employed. These strains, however, did exhibit preferentially high DNA relatedness to a second reference strain.

Antigens, Bacterial↗

Polynucleotide sequence divergence among strains of Escherichia coli and closely related organisms.

Polynucleotide sequence similarity tests were carried out to determine the extent of divergence present in a number of Escherichia coli strains, obtained from diverse human, animal, and laboratory sources, and closely related strains of Shigella, Salmonella, and the Alkalescens-Dispar group. At 60 C, relative reassociation of deoxyribonucleic acid (DNA) from the various strains with E. coli K-12 DNA ranged from 100 to 36%, with the highest level of reassociation found for three strains derived from K-12, and the lowest levels for two "atypical" E. coli strains and S. typhimurium. The change in thermal elution midpoint, which indicates the stability of DNA duplexes, ranged from 0.1 to 14.5 C, with thermal stability closely following the reassociation data. Reassociation experiments performed at 75 C, at which temperature only the more closely related DNA species form stable duplexes, gave similar indications of relatedness. At both temperatures, Alkalescens-Dispar strains showed close relatedness to E. coli, supporting the idea that they should be included in the genus Escherichia. Reciprocal binding experiments with E. coli BB, 02A, and K-12 yielded different reassociation values, suggesting that the genomes of these strains are of different size. The BB genome was calculated to be 9% larger than that of K-12, and that of 02A 9% larger than that of BB. Calculation of genome size for a series of E. coli strains yielded values ranging from 2.29 x 10(9) to 2.97 x 10(9) daltons. E. coli strains and closely related organisms were compared by Adansonian analysis for their relatedness to a hypothetical median strain. E. coli 0128a was the most closely related to this median organism. In general, these data compared well with the data from reassociation experiments among E. coli strains. However, anomalous results were obtained in the cases of Shigella flexneri, S. typhimurium, and "atypical" E. coli strains.

Base Sequence↗

Deoxyribonucleic acid relatedness among species of Erwinia and between Erwinia species and other enterobacteria.

Relatedness in species of Erwinia was assessed by determining the extent of reassociation in heterologous deoxyribonucleic acid preparations. Thermal elution chromatography on hydroxyapatite was used to separate reassociated nucleotide sequences from nonreassociated sequences and to determine the thermal stability of related nucleotide sequences. An apparent 15% core of relatedness is present between fire blight, soft-rot, and "atypical" Erwinia species. All Erwinia species showed low to moderate reaction with representative enteric bacteria.

Biological Evolution↗