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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 39 (1992), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A method for the isolation of micronuclear DNA from Paramecium tetraurelia has been developed. After cell lysis, a low speed centrifugation at 1,000 g is used to remove all of the unbroken cells and macronuclei and approximately two thirds of the macronuclear fragments. Next a higher speed centrifugation of 9,000 g sediments the micronuclei and frees them from small particulates and soluble constituents. Advantage is then taken of the fact that micronuclei have a lower density than do macronuclear fragments in 45%–60% Percoll. Micronuclei float to the top during centrifugation at 24,000 g, while macronuclear fragments sediment. After several cycles of centrifugation in Percoll, the micronuclei, although heavily contaminated with cytoplasmic components, are essentially free of macronuclei and macronuclear fragments. Micronuclear DNA can then be extracted from the suspension. The whole procedure is very rapid and in about an hour micronuclear and macronuclear DNA can be separated. About 2 μg of micronuclear DNA can be obtained from 6 times 107 paramecia. We find that there are internal sequences in the micronuclear A gene DNA in wild type cells which are eliminated when the micronuclei develop into macronuclei. They yield unique restriction fragments for micronuclei and macronuclei. Therefore the purity of the preparations is easily monitored by probing Southern blots of restriction enzyme-digested DNA with the cloned A gene. No differences have been found between the micronuclear A gene in wild type and the d48 mutant.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 26 (1979), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Synopsis.A considerable amount of evidence is now available to indicate that the DNA in the ciliates Oxytricha and Stylonychia undergoes fragmentation when the micronucleus forms a macronucleus. Some evidence suggests that fragmentation may also occur in Tetrahymena and possibly in Paramecium. It is shown that some regulatory or nonrandom segregational mechanism must operate during cell divisions to maintain genic balance in Tetrahymena. Both the hypothesis of macronuclear subunits and also a new hypothesis based on replicative control of DNA are capable of explaining the currently know biochemical, cytological, and genetic facts.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 34 (1987), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Several genes for surface antigens of the Paramecium aurelia complex of species have been isolated. In addition lo known deletions of the 51A gene, we have obtained deletions involving the 51B gene and have developed a procedure for obtaining deletions of additional genes. Both Mendelian and non-Mendelian deletions of both the A and B genes have been found. In the non-Mendelian deletions the genes are present in the micronuclei and absent in the macronuclei. Processing of micronuclear DNA into new macronuclear DNA at conjugation and autogamy is under the control of the old macronucleus, and newly forming macronuclei become exactly like the old. Thus in the non-Mendelian mutants, macronuclei have a specific antigen gene deleted and also are impaired in their ability to direct normal DNA processing at the next conjugation or autogamy. These cases, along with others, show that this system of macronuclear control is a fundamental feature of ciliate genetics. The sequence of the 51A and 51C genes is described and compared with the 156G and 51H genes obtained by others. The 51A and 156G genes are remarkably similar while 51Cand 51H are rather different. No introns or pseudogenes have been observed. Some, possibly all, of the genes are on the ends of chromosomes. Characteristic upstream and downstream sequences adjacent to the coding portions of the genes are given. The sequences UAA and UAG are preferred over CAA and CAG for glutamine while UGA is the true stop codon.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 16 (1969), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: SYNOPSIS. A spiral, rod- or crescent-shaped symbiont here designated alpha, is present in the macronucleus of killer stock 562, syngen 2 of Paramecium aurelia. This stock has a cytoplasmic symbiont, kappa, as well as alpha. Lines were obtained which had only alpha, others which had only kappa, and some which had neither. It was possible to purify and separate both kinds of symbiont from homogenates of stock 562 using an ECTEOLA column. The killing action of this stock is due to kappa, not alpha. Observations on the structure of alpha with the electron microscope indicate that alpha, like the cytoplasmic symbionts in this species, is a bacterium. Alpha is never seen in the micronucleus, is rarely found in the cytoplasm, but abounds in the macronucleus. If paramecia are allowed to grow slowly after autogamy, alpha passes from the old macronuclear fragments, infects the new macronucleus, and all animals retain alpha. In exautogamous paramecia growing at maximum fission rate, however, alpha often does not infect the new macronucleus and is lost from many lines when the old macronuclear fragments disappear. In mixed cultures containing alpha-bearing and alphafree paramecia, it has been found that alpha readily invades the macronucleus of paramecia of susceptible stocks. Homogenates of alpha-bearing cultures are also infective. Infection is highly specific, occurring in only 6 of the 44 stocks of P. aurelia in which infection was attempted, and these 6 are all syngen 2. It is suggested that the short rod or crescent form of alpha is the reproductive form, while the elongated spiral form is probably the invasive motile form.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 111 (1971), S. 202-208 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Phage-like particles from kappa of stock 562 of Paramecium aurelia have been isolated by CsCl density gradient centrifugation. Analyses show that the particles contain about 1.6×1016g DNA and 2.0×10-16g protein. Their buoyant density is approximately 1.47. DNA from the particles has a buoyant density very close to that of whole kappa DNA. The presence of DNA in the particles has been confirmed by a cytochemical technique. The results support the conclusion that kappa contains a bacteriophage.
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