Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors

Author: Yanisch-Perron, C.; Vieira, J.; Messing, J.

Description: 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.

Subject headings: 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

Publication year: 1985

Journal or book title: Gene

Volume: 33

Issue: 1

Pages: 103-119

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Type: Journal Article

Serial number: 274