Isolation and Characterization of Rifampin-Resistant and Streptolydigin-Resistant Mutants of Bacillus subtilis with Altered Sporulation Properties

Mutants of Bacillus subtilis with altered deoxyribonucleic-dependent ribonucleic acid polymerase activity have been isolated and characterized. These mutants, selected as strains resistant to rifampin or streptolydigin, demonstrate drug-resistant in vitro ribonucleic acid synthesis. Sporeforming ability and support of phage infection are altered in many of the mutants. Mutations to rifampin and streptolydigin resistance have been located on the B. subtilis chromosome and ordered relative to the markers cysA14 and str.     

The Distribution of Crossovers along Unreplicated Lambda Bacteriophage Chromosomes

The distribution of crossovers along unreplicated chromosomes of bacteriophage lambda has been examined by determining the density distributions and genotypes of particles in the progenies of crosses of density-labeled by ordinary parents in the presence of genetic blocks to replication. The Red and Rec systems combined produce crossovers primarily near the ends (especially the right end) of the chromosome. Removal of the generalized lambda recombination functions by red and gam mutations results in loss of these terminal crossovers; coupled with this loss is a disappearance of the differential dependence of recombination frequencies in terminal and central intervals on DNA synthesis. Removal of the bacterial system by a recA mutation results in severe depression of crossing over among unreplicated phage, with the few recombinants produced by the lambda system occurring near the right end.     

Evidence for the Involvement of Serine Transhydroxymethylase in Serine and Glycine Interconversions in SALMONELLA TYPHIMURIUM

Salmonella typhimurium can normally use glycine as a serine source to support the growth of serine auxotrophs. This reaction was presumed to occur by the reversible activity of the enzyme, serine transhydroxymethylase (E. C. 2. 1. 2. 1; L-serine: tetrahydrofolic-5, 10 transhydroxymethylase), which is responsible for glycine biosynthesis. However, this enzyme had not been demonstrated to be solely capable of synthesizing serine from glycine in vivo. The isolation and characterization of a mutant able to convert serine to glycine but unable to convert glycine to serine supports the conclusion that a single enzyme is involved in this reversible interconversion of serine and glycine. The mutation conferring this phenotype was mapped with other mutations affecting serine transhydroxymethylase (glyA) and assays demonstrated reduced activities of this enzyme in the mutant.     

Reductive Pentose Cycle and Formate Assimilation in Rhodopseudomonas palustris

Rhodopseudomonas palustris assimilated formate autotrophically as carbon dioxide and hydrogen arising from the activity of the formic hydrogenlyase system. Kinetic analyses of cell suspensions pulse-labeled with (14)C-formate or (14)C-bicarbonate showed similar distributions of incorporated radioactivity. In both cases phosphate esters were the first assimilation products. Ribulose diphosphate carboxylase, phosphoribose isomerase, and phosphoribulokinase, characteristic enzymes of the reductive pentose cycle, were present in extracts of cells grown on formate.     

Methacarn (methanol-Carnoy) fixation

Summary According to chemical data, methanol raises the shrinkage temperature of collagen significantly more than ethanol (86° C versus 70° C). Since increase of shrinkage temperature appears desirable in tissues to be embedded in paraffin, methanol was substituted for ethanol in Carnoy's fluid. This methanol-Carnoy mixture is referred to as methacarn solution. The fixation-embedding procedure was similar to that described in the study of Carnoy fixation. Methacarn-fixed sections showed little or no shrinkage and compared well with material fixed in Carnoy's or Zenker's fluid. Myofibrils, especially in endothelial and epithelial cells, were more prominent in methacarn- than in Carnoy-fixed tissues.A review of the chemical literature showed that methanol, ethanol and chloroform stabilize or even enhance helical conformations of proteins, presumably by strengthening of hydrogen bonds. Interference with hydrophobic bonds causes unfolding and/or structural rearrangements in globular proteins. The twin-helical structure of DNA collapses in alcoholic solutions. Hence, methacarn fixation can be expected to preserve the helical proteins in myofibrils and collagen, but the conformations of globular proteins and DNA will be significantly altered. Literature on conformational effects produced by fixatives used in electron microscopy was also reviewed. Glutaraldehyde and OsO₄ cause considerable loss of helix (22–29% and 39–66% respectively). KMnO₄ and glutaraldehyde followed by OsO₄ produce extensive transitions from helical to random-coil conformations similar to those seen in powerful denaturants such as 8 M urea. Evidently these fixatives are unsuitable for studies of helical proteins. In contrast ethylene glycol preserves helical conformations.     

Differential Tolerance of Streptomycetes to Sodium Chloride as a Taxonomic Aid

A survey was made of the NaCl tolerance of approximately 1,300 Streptomyces strains belonging to 313 species. The growth medium of the organisms was supplemented with a graded series of NaCl concentrations (4, 7, 10, and 13%). Only 1.8% of the species could not tolerate 4% NaCl; 26.9% could grow at a maximum of 4%; 49.7% could tolerate a maximum of 7%; 18.8% could grow at a maximum of 10%; and only 2.8% could tolerate 13% NaCl. In evaluating the relationships of NaCl tolerance to various taxonomic features, higher tolerance was statistically associated with the "yellow" and possibly the "white"-spored streptomycetes, whereas the "red"-spored series tended to have lesser tolerance. Higher tolerance was also indicated for spiny-spored species, as a group, than for smooth-spored forms. Likewise, nonproducers of melanin, collectively, were more NaCl tolerant than melanin-producing species. Uniformity of test responses between strains of species studied suggested the usefulness of NaCl tolerance as a taxonomic criterion.     

On the mechanism of sequence iron-hematein stains

Summary As a prerequisite for the histochemical study of sequence iron-hematoxylin stains the iron alum-acidified hematein procedure was developed which does not require differentiation.Histochemical blocking and extraction procedures demonstrated that carboxyl and hydroxyl groups are essential for the binding of cationic iron.The iron alum-Prussian blue reaction colored collagen, reticulum fibers and basement membranes more intensely than muscle fibers. Treatment of tissue sections mordanted in iron alum with the acidified hematein solvent resulted in practically complete removal of iron from all tissue structures. It must therefore be concluded that the selective staining of muscle fibers, terminal bars and related structures with sequence iron-hematein stains is not due to high affinity of iron for these tissue components.Observations by R. and M. Heidenhain on sequence hematoxylin-potassium dichromate and hematoxylin-alum stains and data from modern textile chemistry indicate that the staining patterns obtained with metal-hematein sequence stains are determined by the affinity of the hematein moiety for certain tissue structures.