Abstract
Newly replicated deoxyribonucleic acid (DNA) in Mycoplasma gallisepticum A5969 is membrane associated. Cells pulse-labeled 1 to 3 min with (3)H-thymidine are lysed by a freeze-thaw procedure. After brief sonic treatment to shear the DNA, differential centrifugation gives a cell fraction (P2) that is enriched sevenfold for pulse-labeled DNA. P2 contains 80% of the total adenosine triphosphatase activity, 65% of the total cholesterol, and morphologically intact terminal bleb structures. Three to four minutes are needed to fully label the DNA growing-point region, whereas 7 to 8 min are required to "chase" 50% of the (3)H-labeled DNA. This pulse-chase removes 80 to 85% of the nascent DNA from the P2 fraction.
MeSH Terms
Adenosine Triphosphatases/metabolism
Bacterial Proteins/analysis
Bacteriological Techniques
Bacteriolysis
Carbon Isotopes
Cell Fractionation
Cell Membrane/analysis,enzymology
Cholesterol/analysis
DNA Replication
DNA, Bacterial/analysis
Isotope Labeling
Microscopy, Electron
Mycoplasma/analysis,cytology,enzymology,metabolism
RNA, Bacterial/analysis
Subcellular Fractions/analysis
Thymidine/metabolism
Tritium
Uridine/metabolism
Vibration
Chemicals
Bacterial Proteins
Carbon Isotopes
DNA, Bacterial
RNA, Bacterial
Tritium
Cholesterol
Adenosine Triphosphatases
Thymidine
Uridine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Quinlan D C
Maniloff J
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9 references, click to expand
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