Nucleic Acids Research, 1981, Vol. 9, No. 13 2999-3014
© 1981
MOLECULAR BIOLOGY |
Molecular cloning of the complete Epstein-Barr virus genome as a set of overlapping restriction endonuclease fragments
Imperial Cancer Research Fund Laboratories, Lincoln's Inn Fields P.O. Box 123, London WC2A 3PX, UK *Department of Clinical Chemistry, University of Gothenburg 413 45 Gothenburg +Department of Medical Biochemistry, University of Gothenburg 400 33 Gothenburg, Sweden
Received May 14, 1981. A complete collection of fragments of Epstein-Barr virus DNA, obtained by cleavage with restriction endonuclease Eco RI, has been cloned. Fourteen different internal fragments of the virus genome, derived from linear virion DNA of the B95-8 strain, and sequences corresponding to the terminal regions of virion DNA, derived from intracel1ular circular EBV DNA isolated from B95-8 cells, were cloned. Sizes of fragments were determined by agarose gel electrophoresis and their sum leads to an estimated molecular weight of 110 x 106 for virion DNA. Large Eco RI DNA fragments of special interest were also cloned in cosmids using another source of EBV DNA, that is, the circular viral DNA derived from Raji cells. In order to provide a set of overlapping sequences, all the 29 internal Bam HI fragments of B95-8 virion DNAwere cloned in pBR322. The map location with in the viral genome of each cloned DNA fragment was identified by hybridizing to blots of virion DNA cleaved with several different restriction endonucleases.
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