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Construction of restriction enzyme fragment libraries containing DNA from human adenovirus types 2 and 5
Authors:A Stenlund  M Perricaudet  P Tiollais  U Pettersson
Institution:1. Department of Microbiology, Biomedical Center, Box 581, S-751 23 UppsalaSweden;7. Recombinaison et expression génétique (Institut National de la Santé et de la Recherche Médical U163) Unité de Génie Génétique, Institut Pasteur, 28, rue du Dr. Roux, 75724 Paris Cedex 15 France
Abstract:Restriction-fragment libraries containing adenovirus type 2 (Ad2) DNA have been constructed, using the pBR322 plasmid (Bolivar et al., 1977) as a vector. Clones have been isolated which contain all the HindIII fragments of Ad2 DNA except the terminal G- and K-fragments inserted into the HindIII cleavage site of the vector. All the 13 SmaI-fragments of Ad2 DNA were separately inserted into the PstI site of the pBR322 vector after addition of homopolymeric poly(dG) tails to the fragments and poly(dC) tails to the linearized plasmid. Two large fragments of adenovirus type 5 (AD5) DNA, located between map positions 17.0 and 59.5 and between map positions 59.5 and 97.3, respectively, were cloned using bacteriophage lambda as a vector. All clones, which are described in the present report, are available upon request.
Keywords:Recombinant DNA  plasmid pBR322  restriction endonucleases  dC-dG tailing  Ad  adenovirus  SDS  sodium dodecyl sulphate
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