Improved M13 Phage Cloning Vectors and Host Strains: Nucleotide Sequences of the M13mp18 and pUC19 Vectors

Gene. 1985;33(1):103-19. doi: 10.1016/0378-1119(85)90120-9.

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 M13mp 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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Cloning, Molecular*
  • Coliphages / genetics*
  • Conjugation, Genetic
  • DNA Restriction Enzymes / metabolism
  • DNA, Single-Stranded / genetics
  • Escherichia coli / metabolism
  • Genetic Vectors*
  • Methylation
  • Mutation
  • Plasmids
  • Recombination, Genetic

Substances

  • DNA, Single-Stranded
  • DNA Restriction Enzymes

Associated data

  • GENBANK/M11454
  • GENBANK/M11662