+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Highly purified micro nuclei and macro nuclei from tetrahymena thermophila isolated by percoll gradients

Highly purified micro nuclei and macro nuclei from tetrahymena thermophila isolated by percoll gradients

Journal of Protozoology 30(1): 21-30

A new procedure is described that utilizes Percoll gradients for purifying micronuclei (MIC) and macronuclei (MAC) from T. therophila. Separation of MIC from MAC during certrifugation in Percoll gradients occurs as a result of their difference in size rather than density. Three kinds of tests were used to evaluate the purity of the nuclei: visualization of the nuclei by light microscopy; examination of the nuclei by EM; and Southern blots of MIC and MAC DNA probed with the 5s rRNA genes or a fragment from the MAC extrachromosomal rDNA molecule. When examined under the light microscope, the isolated MIC and MAC have much lower nuclear cross contamination levels than previous methods have reported. MIC contaminated with < 1 MAC in 1000 MIC and MAC contaminated with < 1 MIC in 500 MAC can be routinely prepared. Quantitative analyses of electron micrographs gave higher estimates of cross contamination in the purified nuclei, which may, in part, be explained by the difficulty in identifying small MIC or MAC fragments. Southern blots of MIC and MAC DNA probed with 5s rDNA confirmed the level of MAC contamination in the MIC estimated by light microscopy during purification of the nuclei. The level of nucleolar contamination in the MIC was estimated at 10% by Southern blots of MIC and MAC DNA, derived from a heterokaryon with distinctive MIC and MAC Bam HI sites, using an rDNA probe.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 005573946

Download citation: RISBibTeXText

Related references

Highly purified lipid X is devoid of immunostimulatory activity. Isolation and characterization of immunostimulating contaminants in a batch of synthetic lipid X. The Journal of Biological Chemistry 265: 59-64, 1990

Histones can provoke immune complex formation in the glomerular capillary wall. Kidney International 33(1): 324, 1988

A comparison of slow release and conventional nitrogenous fertilizers for an established pangola grass pasture in a subtropical environment. Australian Journal of Experimental Agriculture and Animal Husbandry 18(92): 415-422, 1978

Histones and chromatin structure. Ts'O, Paul O. P. (ed.). The Molecular Biology Of The Mammalian Genetic Apparatus, Vol. 1. International Symposium. Pasadena, Calif., Usa, Dec.,. Xx 444p. Illus. Elsevier Excerpta Medica North-Holland: Amsterdam, Netherlands; Elsevier North-Holland, Inc: New York, N.y., Usa. Isbn 0-7204-0625-0. 53-70, 1977

Numbers food consumption and fish predation by birds in lake mockeln southern sweden. Ornis Scandinavica 7(1): 61-70, 1976

Organization of the 5.8s 16 18s and 23 28s ribosomal rna genes of cephalosporium acremonium. Current Genetics 12(3): 209-214, 1987

Pattern of hindlimb malformations produced by dominant hemimelia dh in dh positive mice. Teratology 33(3): 41C-42C, 1986

Studies on histone fraction f 2a 1 in macro nuclei and micro nuclei of tetrahymena pyriformis. Journal of Cell Biology 57(3): 773-781, 1973

Numbers biomass and productivity of microorganisms in the soil. Kosmos Seria A Biologia 22(4): 379-387, 1973

Electron microscopic study of nucleo cytoplasmic interrelationships in ecto placental cone. Anatomical Record 169(2): 391, 1971

Isolation of micro fibrils from rat vibrissal follicles. Journal of Cell Biology 52(1): 174-179, 1972

Polymorphism of micro nuclei of paramecium caudatum part 4 comparative analysis of nuclear reorganization during progamous divisions of 4 morphological types. Tsitologiya 13(10): 1289-1298, 1971

Timing of termination viewed from nonstress test in high risk pregnancies. Acta Obstetrica et Gynaecologica Japonica (Japanese Edition) 36(11): 2512-2513, 1984

Micro nuclei induced by x rays and chemical mutagens in meiotic pollen mother cells of tradescantia paludosa a promising mutagen test system. Mutation Research 64(5): 307-314, 1979

Histones of chick erythrocytes fractioning of total histones and isolation of a specific histone. 1968