Vicilin genes ofPisum

Summary We describe four genomic clones of pea 7S storage protein gene, one of which corresponds to convicilin, and the others to vicilin. Hybridization studies exploiting these clones, and previously identified cDNA clones, have enabled us to define six different loci. Three of these loci have been mapped to positions on chromosome 7.     

Component X. An immunologically distinct polypeptide associated with mammalian pyruvate dehydrogenase multi-enzyme complex

The mammalian pyruvate dehydrogenase multi-enzyme complex contains a tightly-associated 50 000-Mr polypeptide of unknown function (component X) in addition to its three constituent enzymes, pyruvate dehydrogenase (E1), lipoate acetyltransferase (E2) and lipoamide dehydrogenase (E3) which are jointly responsible for production of CoASAc and NADH. The presence of component X is apparent on sodium dodecyl sulphate/polyacrylamide gel analysis of the complex, performed in Tris-glycine buffers although it co-migrates with the E3 subunit on standard phosphate gels run under denaturing conditions. Refined immunological techniques, employing subunit-specific antisera to individual components of the pyruvate dehydrogenase complex, have demonstrated that protein X is not a proteolytic fragment of E2 (or E3) as suggested previously. In addition, anti-X serum elicits no cross-reaction with either subunit of the intrinsic kinase of the pyruvate dehydrogenase complex. Immune-blotting analysis of SDS extracts of bovine, rat and pig cell lines and derived subcellular fractions have indicated that protein X is a normal cellular component with a specific mitochondrial location. It remains tightly-associated with the 'core' enzyme, E2, on dissociation of the complex at pH 9.5 or by treatment with 0.25 M MgCl2. This polypeptide is not released to any significant extent from E2 by p-hydroxymercuriphenyl sulphonate, a reagent which promotes dissociation of the specific kinase of the complex from the 'core' enzyme. Incubation of the complex with [2-14C]pyruvate in the absence of CoASH promotes the incorporation of radio-label, probably in the form of acetyl groups, into both E2 and component X.     

Changes in density of DNA after interaction with dipicolinic acid and its possible role in spore heat resistance

We examined the effect of interacting dipicolinic acid and its calcium chelate on the wet and dry density of DNA. Complexes are produced whose densities are different from those of the individual components. Also, we observed two modes of binding, one strong the other weak, between DPA or CaDPA and DNA. The strength of the binding modes was reflected in the rate of dissolution of the complexes as monitored by changes in wet density with time and temperature. We conclude from these and other data in the literature that the interaction of dipicolinic acid with DNA not only influences the spore wet density and the ratio of core/core⁺ cortex volume, but may also influence the spore heat resistance.     

A NEW GENUS OF PROTOSTELIDS SHOWING AFFINITIES WITH CERATIOMYXA

A new genus and species of the Protosteliida (Mycetozoa), Ceratiomyxella tahitiensis, was isolated from dead plant material—var. tahitiensis from Tahiti and var. neotropicalis from Brazil and Colombia. The sporocarps have deciduous spores borne singly on slender hollow stalks; zoocysts with anteriorly flagellate planonts are produced. The trophic stage is comprised of uninucleate to plurinucleate amoeboid cells and reticulate plasmodia; the uninucleate cells become flagellate in water. The prespore cells and spores are plurinucleate. Sexuality has not been demonstrated. Var. tahitiensis has globose spores and produces its zoocysts just after spore germination, whereas var. neotropicalis has subglobose spores and forms zoocysts later in the life cycle. The species is thought to show phylogenetic relationships with Ceratiomyxa, which was recently transferred to the Protosteliida by Olive.     

Internal proteins of bacteriophage T4D: Their characterization and relation to head structure and assembly

Three basic proteins of low molecular weight (about 8000, 10,000 and 18,000) were isolated from the T4D phage particle. Many molecules of each protein are located within the phage head, possibly in association with the DNA, and together with the proteins which form the head membrane comprise most of the head structural protein. The purified internal proteins were characterized by physicochemical and immunological techniques; a radio-immunoassay allowed measurement of their synthesis in phage infected bacteria. Each internal protein is synthesized at both early and late times after infection. Their structural genes are present in the phage genome, but do not appear to be among the known amber mutant-containing genes of T4D. No evidence was found to suggest that the internal proteins are formed from a common precursor molecule, nor are their origins related to those of the internal peptides; however, one of the internal proteins may be altered before its incorporation into the phage. Pulse-chase experiments with two of these proteins show that they are incorporated into certain defective T4D heads. Whether or not they are incorporated appears to depend on the degree of completion of these heads, perhaps with respect to DNA packaging.     

Effect of high temperature on infectivity of Toxoplasma gondii tissue cysts in pork

To study the effect of high temperature on infectivity of Toxoplasma gondii tissue cysts, pork from infected pigs was mixed with infected mouse brains and homogenized thoroughly. Twenty-gram samples of infected homogenized meat were sealed in plastic pouches, pressed to a uniform thickness of 2 mm, and subjected to water-bath temperatures of 49, 52, 55, 58, 61, 64, and 67 C for 0.01, 3, 6, 9, 12, 24, 48, and 96 min. Treated samples were digested in HCl-pepsin solution and bioassayed in mice. Toxoplasma gondii tissue cysts remained viable at 52 C for 9.5 min but not for 9.5 min at 58 C; tissue cysts were generally rendered nonviable by heating to 61 C or higher temperature for 3.6 min. Tissue cysts survived once at 64 C for 3 min. These data demonstrate that T. gondii tissue cysts are less heat resistant than encysted Trichinella spiralis larvae.     

Evaluation of anti-coccidial drugs' inhibition of Neospora caninum development in cell cultures

Eight anti-coccidial drugs were examined for their efficacies in preventing development of Neospora caninum in bovine monocyte cell cultures. Lasalocid sodium (0.05 microgram/ml), monensin sodium (0.05 microgram/ml), piritrexim (0.01 microgram/ml), pyrimethamine (0.05 microgram/ml), and trimethoprim (5.0 micrograms/ml) were effective in preventing development of intracellular N. caninum tachyzoites (P less than 0.05). No differences (P greater than 0.05) in mean numbers of infected cells compared to controls were observed in cultures treated with amprolium hydrochloride (10.0 micrograms/ml), sulfadiazine (200.0 micrograms/ml), and sulfamethoxazole (200.0 micrograms/ml).