An improved filamentous helper phage for generating single-stranded plasmid DNA

Marjorie Russel, Simon Kidd, Mark R. Kelley

Research output: Contribution to journalArticle

240 Scopus citations

Abstract

Gene cloning in plasmid vectors that contain a filamentous phage intergenic region presents several advantages. However, technical difficulties have been a problem, primarily low yields of packaged single stranded (ss) plasmid DNA from the rapid, small scale procedures usually employed, and ambiguities in sequencing reactions attributed to the contamination by helper phage ss DNA. We report here the construction and some properties of a new f1 helper phage. Using this phage, R408, plasmid ss DNA is packaged and exported preferentially over phage ss DNA, and the absolute yield of plasmid ss DNA is usually increased.

Original languageEnglish (US)
Pages (from-to)333-338
Number of pages6
JournalGene
Volume45
Issue number3
DOIs
StatePublished - 1986
Externally publishedYes

Keywords

  • Recombinant DNA
  • bacteriophage f1
  • deletions
  • gene cloning
  • packaging signal

ASJC Scopus subject areas

  • Genetics

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