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.
Keywords
pUC19BiologyCloning vectorCloning (programming)Multiple cloning siteGeneticsPlasmidVector (molecular biology)Molecular cloningNucleic acid sequenceGeneMolecular biologyComputational biologyRecombinant DNAComplementary DNA
MeSH Terms
Base SequenceCloningMolecularColiphagesConjugationGeneticDNA Restriction EnzymesDNASingle-StrandedEscherichia coliGenetic VectorsMethylationMutationPlasmidsRecombinationGenetic
Affiliated Institutions
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Publication Info
- Year
- 1985
- Type
- article
- Volume
- 33
- Issue
- 1
- Pages
- 103-119
- Citations
- 15137
- Access
- Closed
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Cite This
Celeste Yanisch-Perron,
Jeffrey Vieira,
Joachim Messing
(1985).
Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors.
Gene
, 33
(1)
, 103-119.
https://doi.org/10.1016/0378-1119(85)90120-9
Identifiers
- DOI
- 10.1016/0378-1119(85)90120-9
- PMID
- 2985470