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Specimens For Observing Single Events Of Dna Metabolism By Fluorescence Microscopy

Published online by Cambridge University Press:  02 July 2020

M. Sun  and
P. Serwer
M. Sun
Affiliation:
Department of Biochemistry, The University of Texas Health Science Center, San Antonio, Texas78284-7760.
P. Serwer
Affiliation:
Department of Biochemistry, The University of Texas Health Science Center, San Antonio, Texas78284-7760.
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Extract

Single stained DNA molecules can be visualized by fluorescence microscopy (1). Therefore, perhaps single events of DNA metabolism can also be visualized by fluorescence microscopy. Problems to solve include (a) photobleaching of stained DNA, (b) photolytic damage to macromolecules necessary for metabolism, including DNA, (c) inhibition of metabolism by the process of preparation of the specimen, and (d) interference with image formation by macromolecules that either must be or by chance are present during metabolism. These problems have been solved, at least in part, during a study of the in vitro packaging of bacteriophage T7 (2). Packaging occurs in T7 capsids that had previously been assembled without assistance of DNA. The DNA substrate for packaging is a head-to-tail multimer (concatemer) of the terminally repetitious, nonpermuted T7 DNA. The metabolic event observed in ref. 2 was capsid-dependent specific cleavage of concatemers during packaging. The concatemers had formed a DNA network before cleavage.

Type
Biological Specimen Preparation
Information
Microscopy and Microanalysis , Volume 5 , Issue S2: Proceedings: Microscopy & Microanalysis '99, Microscopy Society of America 57th Annual Meeting, Microbeam Analysis Society 33rd Annual Meeting, Portland, Oregon August 1-5, 1999 , August 1999 , pp. 1312 - 1313
Copyright
Copyright © Microscopy Society of America

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References

(1)Bustamante, C., Annu. Rev. Biophys. Biophys. Chem. 20 (1991) 415.CrossRefGoogle Scholar
(2)Sun, M., et.al., Biochemistry 36, (1997) 13018.CrossRefGoogle Scholar
(3)Serwer, P., et. al., Biophys. J. 69, (1995) 2649.CrossRefGoogle Scholar