Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors

Celeste Yanisch-Perron, Jeffrey Vieira, Joachim Messing

Research output: Contribution to journalArticlepeer-review

11456 Scopus citations

Abstract

Three kinds of improvements have been introduced into the M13-based cloning systems. (1) New Escherichia coli host strains have been constructed for the E. coli bacteriophage M13 and the high-copy-number pUC-plasmid cloning vectors. Mutations introduced into these strains improve cloning of unmodified DNA and of repetitive sequences. A new suppressorless strain facilitates the cloning of selected recombinants. (2) The complete nucleotide sequences of the M 13mp and pUC vectors have been compiled from a number of sources, including the sequencing of selected segments. The M13mp18 sequence is revised to include the G-to-T substitution in its gene II at position 6 125 bp (in M13) or 6967 bp in M13mp18. (3) M13 clones suitable for sequencing have been obtained by a new method of generating unidirectional progressive deletions from the polycloning site using exonucleases HI and VII.

Original languageEnglish (US)
Pages (from-to)103-119
Number of pages17
JournalGene
Volume33
Issue number1
DOIs
StatePublished - 1985

All Science Journal Classification (ASJC) codes

  • Genetics

Keywords

  • Recombinant DNA
  • molecular cloning
  • polycloning sites
  • progressive deletions

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