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W R Chesbro

Publications and source records attributed to W R Chesbro.

9 recordsLinked to original sources
PubMed1994

Electro-transformation of Clostridium beijerinckii NRRL B-592 with shuttle plasmid pHR106 and recombinant derivatives.

Conditions for transformation of the solventogenic anaerobe Clostridium beijerinckii NRRL B-592 with plasmid DNA via electroporation are described. Shuttle plasmid pHR106 and two derivatives constructed in this study were transferred and were expressed in this organism. One recombinant derivative of pHR106 was constructed by separately subcloning the clostridial tetracycline (tetP) resistance genes into pHR106. The second vector conferring erythromycin resistance was obtained via in-vivo recombination. The new constructs, termed pRZL and pRZE respectively, were then transferred to C. beijerinckii in order to evaluate their potential as shuttle vectors. The recombinant plasmids were shown to transfer to C. beijerinckii and were expressed as autonomously replicating vectors. The use of these plasmids as cloning and shuttle vectors for C. beijerinckii is discussed.

Chromosomes, Bacterial↗
PubMed1994

Improved techniques for dissociating particle-associated mixed ruminal microorganisms from ruminal digesta solids.

Combinations of physical and chemical methods were evaluated for their ability to remove particle-associated microorganisms (PAM) from saline-washed ruminal digesta solids (SWRDS). Physical methods included chilling and storage, homogenization, multiple extraction, and agitation with marbles. Chemical methods included use of low pH, Tween 80, formaldehyde, methanol, tertiary butanol, and methylcellulose. Microbial removal from SWRDS was determined directly by using epifluorescence microscopy and indirectly by measuring removal of diaminopimelic acid and total purines. Different combinations of methods resulted in removals of 46 to 82% for particle-associated bacteria (PAB), 52 to 98% for particle-associated protozoa (PAP), and 60 to 83% for PAB plus PAP. Two methods were considered most effective, based on microscopy; both removed similar amounts of PAB (79 to 82%) and PAB plus PAP (80 to 83%). In one method, SWRDS were stored for 24 h at 4 degrees C in a solution of pH 2 saline, .1% Tween 80, 1.0% methanol, and 1.0% tertiary butanol. In the other method, SWRDS were incubated for 30 min in .1% methylcellulose before storage for 24 h at 4 degrees C in pH 2 saline, .1% Tween 80, and 1.0% methanol. Common to both treatments was subsequent homogenization of the suspensions for 15 s followed by washing the digesta solids seven times with the treatment solutions. Both methods resulted in values that exceeded those reported previously for removal of PAM from ruminal digesta solids.

Animal Feed↗
PubMed1985

Integration of the Vwa plasmid into the chromosome of Yersinia pestis strains harboring F' plasmids of Escherichia coli.

Conditional virulent strains of Yersinia pestis were used as recipients of an F-lac plasmid from either Escherichia coli 23.10S or E. coli CSH23. The transconjugants of Y. pestis were calcium independent; however, calcium dependence was restored after the loss of the E. coli plasmid. The plasmid contents of several Y. pestis F-lac clones were compared with those of the parent strains and several lac segregants. Although the 6- and 61-megadalton plasmids were detected in all strains tested, the Vwa plasmid was absent in the F-containing strains. The plasmid was evident, however, in the lac segregants derived from the F-lac clones. Results of Southern transfers indicated that the Vwa plasmid had integrated into the chromosome of Y. pestis strains harboring F-lac.

Calcium↗
PubMed1984

Eubacteria have 3 growth modes keyed to nutrient flow. Consequences for the concept of maintenance and maximal growth yield.

Aerobic growth of Escherichia coli and Paracoccus denitrificans has been studied in chemostat, fed batch, and recycling fermentor modes under carbon and energy limitation. Two abrupt drops or discontinuities in molar growth yield, Y, have been found that occur over relatively short ranges in the value of specific growth rate. Before the first discontinuity, Y is constant and maximal. After the first discontinuity, at a doubling time of 33 h, Y becomes constant again and independent of mu until the second discontinuity appears at a doubling time of about 50 h, corresponding to a mu of about 0.014. At this point, Y drops to a lower value that is constant at doubling times longer than 100 h, corresponding to a mu of about 0.007. The second discontinuity is associated in Paracoccus with elevated levels of guanosine tetraphosphate (ppGpp) that impose stringent regulation as has been found previously with Bacillus and Escherichia species. It is thus likely that the stringent response generally occurs in bacteria in vivo at a doubling time of about 50 h. The cause of the first discontinuity is unknown. All experiments indicate that Pirt-type calculations relating mu, Y, and maintenance energy demand are no longer valid. In chemostat experiments, the intercept of the relationship between specific substrate utilization and specific growth rate is defined as maintenance. However, this intercept most probably is caused by stringent regulation at low dilution rates. Three regions of bacterial growth rates are defined by this study, corresponding to doubling times of 0.5 to 15 h, 33 to 50 h, and greater than 100 h. Some growth behavior in each region is unique to that region.

Chromatography, Thin Layer↗
PubMed1968

Characteristics of secretion of penicillinase, alkaline phosphatase, and nuclease by Bacillus species.

The distribution of alkaline phosphatase and nuclease activity between cells and medium was examined in one strain of Bacillus licheniformis and four strains of B. subtilis. Over 95% of both activities was found in the medium of the B. licheniformis culture, but in the B. subtilis cultures the amount of enzyme activity found in the medium varied with the strain and the enzyme considered. B. licheniformis 749 and its penicillinase magnoconstitutive mutant 749/C were grown in continuous culture with phosphorous as the growth-limiting factor, and the kinetics of penicillinase formation and secretion were examined. Nutrient arrest halted secretion (usually after a lag of about 30 min) in both the inducible and constitutive strains. Chloramphenicol did not eliminate secretion, but under certain circumstances reduced its rate. In the inducible strain treated with a low level of inducer, the rate of secretion was more affected by the rate of synthesis than by the level of cell-bound enzyme. During induction, the onset of accretion of cell-bound penicillinase and secretion of the exoenzyme were nearly simultaneous. It seems unlikely that a long-lived, membrane- or cell-bound intermediate is mandatory in the secretion of the three enzymes by Bacillus species. In the case of penicillinase secretion, there are at least two different phases. When penicillinase synthesis is proceeding rapidly, the rate of secretion is five to six times greater at equivalent concentrations of membrane-bound penicillinase than it is when penicillinase synthesis is reduced. The data require that any membrane-bound intermediate in the formation of exoenzyme be much shorter-lived in cells with a high rate of synthesis than in cells with a low rate. Either there are two separate routes for the secretion of penicillinase or the characteristics of the process vary substantially between the early stages and the declining phase of induction.

Alkaline Phosphatase↗
PubMed1967

Enzymatic detection of the growth of Staphylococcus aureus in foods.

A specific method has been developed for the extraction and measurement of staphylococcal nuclease in foods in which Staphylococcus aureus has grown. The method was used to compare staphylococcal growth with nuclease production in foods under varying conditions of temperature, aerobiosis, and competition from other microorganisms. It was concluded that the nuclease is produced under any conditions that permit growth of S. aureus, and little or no interference with the test was encountered either from mixed, natural populations or from a variety of pure, laboratory cultures. Nuclease and enterotoxin A production were shown to vary in synchrony for the 234 (Casman) strain of S. aureus, and the sensitivity of the enzymatic detection of nuclease was comparable to the sensitivity of serological detection of enterotoxin A. It was found that 15 min at 121 C was required to reduce the nuclease activity in slurries of contaminated ham below the level present in the unheated slurry. The extraordinary heat resistance of the nuclease permits its detection even in foods heated subsequent to the growth of S. aureus. The nuclease analysis requires about 3 hr to complete and requires no unusual equipment or reagents.

Deoxyribonucleases↗