The Admid System: Generation of Recombinant Adenoviruses by Tn7-Mediated Transposition in

A new system has been developed for generating recombinant adenoviruses by Tn7-mediated transposition in . Low copy number plasmids containing a full-length adenoviral genome with Z -Tn7 replacing E1 have been constructed. The adenovirus plasmid or admid, as well as high copy number progenitors, wer...

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Bibliographic Details
Published inBioTechniques Vol. 29; no. 1; pp. 146 - 154
Main Authors Richards, Cynthia A, Brown, Chadwick E, Cogswell, John P, Weiner, Michael P
Format Journal Article
LanguageEnglish
Published Future Science Ltd 01.07.2000
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Summary:A new system has been developed for generating recombinant adenoviruses by Tn7-mediated transposition in . Low copy number plasmids containing a full-length adenoviral genome with Z -Tn7 replacing E1 have been constructed. The adenovirus plasmid or admid, as well as high copy number progenitors, were stably maintained in strain DH10B. Several transfer vectors containing a mammalian expression cassette flanked by Tn7R and Tn7L were used as donors to transpose the mini-Tn7 into the E1 region of the adenoviral genome. Transposed recombinant admids are readily identified by their β-galactosidase phenotype. Transfection of admid DNA into producer cells resulted in the efficient production of infectious adenovirus. This easy-to-use, efficient system generates pure, clonal stocks of recombinant adenovirus without successive rounds of plaque purification.
ISSN:0736-6205
1940-9818
DOI:10.2144/00291rr01