[Isolation and various properties of the chromosome gene coding the frog ro-crystallin].
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Biomedical subjects
Publications and source records attributed to G G Gauze.
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Sibiromycin added to linear chromosomal E. coli DNA in vitro leads to the decrease of bouyant density in neutral CsCl density gradient. This decrease is a linear function of sibiromycin/DNA ratio and amounts to about 32 mg/ml at the ratio equal to 0.1. Binding sibiromycin does not change the degree of hydration of DNA as revealed by centrifugation in metrizamide density gradients. When added to the covalently closed or open circular DNA of PM-2 phage, sibiromycin decreased the bouyant density of these DNA species to a similiar extent. The antibiotic does not induce single-strand breaks in DNA in vitro as follows from the results of ethidium bromide-CsCl density gradient centrifugation of covalently closed PM-2 DNA.
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A cytological hybridization of H-3-complementary RNA synthetized from DNA a template of D. virilis with the polytene chromosomes of D. virilis and the hybrids between D. virilis and D. texana, was carried out in situ. The uridine label of RNA was shown to be located mainly over the disc of the polytene chromosomes, the silver grains in interspecies hybrids being located over both homogous chromosomes including the unpaired gions.
Nuclei isolated from developing embryos of the silkworm (Bombyx mori) are capable of RNA synthesis in vitro. The incorporation of C-14-uridine triphosphate (C-14-UTP) into the acid-insoluble fraction requires the presence of the other three ribonucleoside triphosphates; it is inhibited by actinomycin, olivomycin, and ethidium bromide, and is stimulated by ammonium sulfate. In a medium of moderate ionic strength C-14-UTP incorporation is slightly inhibited by alpha-amanitine (4 mug/ml), whereas in the presence of 0.2 M ammonium sulfate the same alpha-amanitine concentration reduces the incorporation by 90%. These data indicate that silkworm embryo nuclei, like nuclei from several other sources, contain two different RNA-polymerase fractions.
Microsomes isolated from the fibroin region of the Bombyx mori silkgland synthesize in the cell-free system a glycin rich polypeptide or polypeptides presumably representing fibroin precursors. Besides microsomes the system requires ATP and ATP-generating system, GTP, soluble protein fraction and tRNA, glycine incorporation is inhibited by puromycin and cycloheximide. It is shown that the synthesis of a polypeptide with high Gly/Lys ratio requires soluble protein fraction isolated from the silk gland at the end of the instar V. When the soluble protein fraction from the larvoe at the early instar V is used the Gly/Lys ratio in the product is markedly lower. These results permit to suggest that fibroin synthesis may be regulated at the level of tRNA aminoacylation.
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A simple procedure for the isolation of animal mitochondrial DNA is described. It includes deproteinization of mitochondrial lysates with phenol, precipitation of total nucleic acids with polyethylene glycol, RNA precipitation by saet and final purification of DNA using gel chromatography on a Sepharose 4B column. MtDNA preparations obtained were free from protein and RNA and were represented predimonantly by the covalently closed molecules. The content of D-looped molecules in these preparations was equal to 45%.