Human adenovirus cloning vectors based on infectious bacterial plasmids

Goutam Ghosh-Choudhury, Yousef Haj-Ahmad, Pamela Brinkley, John Rudy, Frank L. Graham

Research output: Contribution to journalArticle

55 Scopus citations

Abstract

By making use of the fact that human adenovirus DNA circularizes in infected cells, and that circular forms of the viral genome are infectious, we have developed an improved adenovirus-based cloning system. A deletion mutant of adenovirus type 5 (Ad5) with deletions in early regions 1 (E1) and 3 (E3) was converted to a bacterial plasmid which can regenerate infectious virus following transfection into human 293 cells. A single XbaI recognition site in the deleted E3 region serves as a site for the insertion of foreign DNA. We have used this system to clone a number of genes into the Ad5 genome and describe the insertion of the neomycin/G418 resistance marker into Ad5 as an example.

Original languageEnglish (US)
Pages (from-to)161-171
Number of pages11
JournalGene
Volume50
Issue number1-3
DOIs
StatePublished - 1986
Externally publishedYes

Keywords

  • Ad5 insertion mutants
  • DNA transfection
  • Recombinant DNA
  • bacteriophage λ cos site
  • gene transfer
  • neomycin resistance
  • shuttle vectors

ASJC Scopus subject areas

  • Genetics

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