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Grouping of Adenoviruses and Identification of Restriction Endonuclease Cleavage Patterns of Adenovirus DNAs Using Infected Cell DNA: Simple and Practical Methods
Author(s) -
Suzuki Nobutaka,
Ueno Tetsuji,
Yamashita Toshiharu,
Fujinaga Kei
Publication year - 1981
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1981.tb00138.x
Subject(s) - restriction enzyme , biology , microbiology and biotechnology , dna , ethidium bromide , endonuclease , cleavage (geology) , nucleic acid thermodynamics , dot blot , southern blot , virology , genetics , base sequence , paleontology , fracture (geology)
Simple and practical methods for grouping of adenoviruses and for identification of restriction endonuclease cleavage patterns of viral DNA were established by using infected cell DNA. DNA homology groupings of adenoviruses could be examined by spot hybridization, and restriction endonuclease cleavage patterns of viral DNAs could be obtained by Southern blot hybridization, by using infected cell DNA. The method was very sensitive and allowed the identification of the cleavage pattern of viral DNA of the inoculum by means of cell DNA extracted from infected cells with undetectable cytopathic effect (CPE). In ethidium bromide‐stained gels without Southern blot hybridization, the restriction endonuclease cleavage pattern of viral DNA could be detected precisely in spite of background staining due to cellular DNA. The preparation of infected cell DNA used in these procedures was technically much easier than that of viral DNA. These methods require only a small number of infected cells and allow many isolates to be investigated with ease.