Analysis of Paramecium macronuclear DNA using pulsed field gel electrophoresis

We have analyzed the macronuclear DNA of Paramecium tetraurelia using orthogonal-field-alternation gel electrophoresis. The mean size of the linear macronuclear DNA molecules is approximately 450 kb. Less than 6% of the macronuclear DNA is larger than 800 kb. Using pulse times of 20, 40, 60 and 90 s...

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Bibliographic Details
Published inThe Journal of protozoology Vol. 36; no. 4; p. 402
Main Authors Phan, H L, Forney, J, Blackburn, E H
Format Journal Article
LanguageEnglish
Published United States 01.07.1989
Subjects
Animals
Blotting, Southern
DNA, Circular - analysis
Electrophoresis, Agar Gel - methods
Nucleic Acid Conformation
Nucleic Acid Hybridization
Paramecium - genetics
Paramecium - metabolism
RNA, Ribosomal - genetics
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Summary:We have analyzed the macronuclear DNA of Paramecium tetraurelia using orthogonal-field-alternation gel electrophoresis. The mean size of the linear macronuclear DNA molecules is approximately 450 kb. Less than 6% of the macronuclear DNA is larger than 800 kb. Using pulse times of 20, 40, 60 and 90 s we show that the macronuclear fragment containing the A type variable surface antigen gene migrates reproducibly as a 320-kb linear DNA. Over the same pulse times we describe the unusual migration of the ribosomal RNA gene (rDNA) of P. tetraurelia. At pulse times of 20 and 40 s the rDNA migrates at limit mobility (300 and 500 kb, respectively) whereas with 60- and 90-s pulse times, 2 components of rDNA are observed; 1 fraction independent of pulse time migrating at limit mobility, and a 2nd component migrating between 100-kb and 400-kb linear markers. Based upon previous electron micrographic studies of Paramecium rDNA as well as data presented here we conclude that the majority of Paramecium rDNA molecules are a circular DNA form.
ISSN:0022-3921
DOI:10.1111/j.1550-7408.1989.tb05535.x