Extrachromosomal Elements in Group N Streptococci

The deoxyribonucleic acid (DNA) of Streptococcus lactis C2, S. cremoris B(1), and S. diacetilactis 18-16 was labeled by growing cells in Trypticase soy broth containing (3)H-labeled thymine. The cells were gently lysed with lysozyme, ethylenediaminetetraacetic acid, and sodium lauryl sulfate. The chromosomal DNA was separated from plasmid DNA by precipitation with 1.0 M sodium chloride. The existence of covalently closed circular DNA in the three organisms was shown by cesium chloride-ethidium bromide equilibrium density gradient centrifugation of the cleared lysate material. In an attempt to correlate the loss of lactose metabolism with the loss of plasmid DNA, lactose-negative mutants of these organisms were examined for the presence of extrachromosomal particles. Covalently closed circular DNA was detected in the lactose-negative mutants of S. lactis C2 and S. diacetilactis 18-16. In S. cremoris B(1), however, no covalently closed circular DNA was observed by using cesium chloride-ethidium bromide gradients. Electron micrographs of the satellite band material from S. lactis C2 and its lactose-negative mutant confirmed the presence of plasmid DNA. Three distinct plasmids having approximate molecular weights of 1.3 x 10(6), 2.1 x 10(6), and 5.1 x 10(6) were observed in both organisms.     

Differentiation studies on separated rabbit early erythroid cells

Haemoglobin-containing cells were removed from cell suspensions of adult rabbit bone marrow by immune lysis, and the remaining cells were layered into BSA density gradients. The top fractions contained early erythroid cells, while fractions near the bottom of the gradient contained granulocytes. Two populations of erythroid cells from anaemic rabbits were resolved by the gradient which differed in their time of maximum stimulation of haem synthesis, in culture with erythropoietin. In addition, a difference in requirement for the presence of erythropoietin in the culture medium was found in separated erythroid cells from rabbits with varying degrees of anaemia.     

Capsella embryogenesis: The suspensor and the basal cell

Summary The suspensor and basal cell ofCapsella were examined with the electron microscope and analyzed by histochemical procedures. The suspensor cells are more vacuolate and contain more ER and dictyosomes, but fewer ribosomes and stain less intensely for protein and nucleic acids than the cells of the embryo. The end walls of the suspensor cells contain numerous plasmodesmata but there are no plasmodesmata in the walls separating the suspensor from the embryo sac. The lower suspensor cells fuse with the embryo sac wall and the lateral walls of the lower and middle suspensor cells produce finger-like projections into the endosperm. At the heart stage the suspensor cells begin to degenerate and gradually lose their ability to stain for protein and nucleic acids.The basal cell is highly vacuolate and enlarges to a size of 150 X 70. An extensive network of wall projections develops on the micropylar end wall and adjacent lateral wall. The nucleus becomes deeply lobed and suspended in a strand of cytoplasm traversing the large vacuole. The cytoplasmic matrix darkens at the late globular stage and histochemical staining for protein becomes very intense. The basal cell remains active after the suspensor cytoplasm has degenerated. It is proposed that the suspensor and basal cell function as an embryonic root in the absorption and translocation of nutriments from the integuments to the developing embryo.Research supported by NSF grant GB 3460 and NIH grant 5-RO 1-CA-03656-09.     

2-Deoxyribose Gene-Enzyme Complex in Salmonella typhimurium: Regulation of Phosphodeoxyribomutase

Phosphodeoxyribomutase, the enzyme which catalyzes the interconversion of 2-deoxyribose-1-phosphate to 2-deoxyribose-5-phosphate, has been partially purified from Salmonella typhimurium. The enzyme had an absolute requirement for manganese ion and was stimulated by glucose-1, 6-diphosphate. Phosphodeoxyribomutase was induced by deoxyribose-5-phosphate and was coordinately regulated with the enzymes thymidine phosphorylase and deoxyribose-5-phosphate aldolase, type II. Mutants deficient in these three enzymes were isolated and mapped close to the threonine locus in S. typhimurium. The three enzymes thymidine phosphorylase, deoxyribose-5-phosphate aldolase, type II, and phosphodeoxyribomutase are controlled by a series of linked genes and appear to constitute an operon.     

Biosynthesis of D-mannitol from D-glucose by Aspergillus candidus

Mannitol has long been known as a product of glucose metabolism by some strains of Aspergillus. Apparently no concerted effort, has been made to develop a practical fermentation process to make mannitol. Work at the Northern Laboratory has shown that nearly all strains of white Aspergillus produce significant amounts of mannitol; many strains of black Aspergillus also have this characteristic. Aspergillus candidus NRRL 305 is an exceptionally good mannitol producer. Studies on a fermentation process were conducted in 20-1, stainless steel fermentors, without baffles. Czapek-Dox medium, modified by addition of corn meal, yeast extract, and enzymatically hydrolyzed casein was the most satisfactory medium tested. Suitable increments of glucose were fed daily to the fermentors. The duration of the fermentation was from 10 to 16 days. The effects of agitation, aeration, temperature, and pH of the medium were studied. Under optimal conditions yields of mannitol approached 50% of the glucose consumed.     

The gas–liquid chromatography of carboxylic acid esters of the urinary 11-deoxy-17-oxo steroids: Determination as n-butyrates

1. The gas-liquid-chromatographic separations of the acetate, propionate, n-butyrate, isobutyrate and n-valerate esters of androsterone, aetiocholanolone and dehydroepiandrosterone were studied on a 1% neopentyl glycol sebacate column. The n-butyrate, isobutyrate and n-valerate esters were well resolved. 2. The three steroids derived from hydrolysed urinary 17-oxo steroid conjugate extracts were analysed by gas-liquid chromatography after conversion into their n-butyrate esters. The results were compared with independent determinations involving chromatography on alumina.