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PMID: 3001643 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

EcoK selection vectors for shotgun cloning into M13 and deletion mutagenesis.

Nucleic acids research ·Vol. 13 ·No. 23 ·1985-12-09 ·Pages 8561-71

Waye MM, Verhoeyen ME, Jones PT, Winter G

Abstract

For shotgun cloning into M13 vectors, a double-stranded cassette of synthetic oligonucleotides containing a SmaI site within the two halves of an EcoK site, has been introduced into the vector M13mp8. Cloning of blunt end DNA into the SmaI site destroys the EcoK site, and recombinants are therefore preferentially selected on transfection into a K strain of E.coli. For deletion mutagenesis using synthetic oligonucleotides, an M13 vector with four copies of the EcoK cassette has been made to facilitate the joining of lacZ or a Factor Xa cleavage site to any protein reading frame.

MeSH Terms

Base Sequence Chromosome Deletion Cloning, Molecular/methods DNA Restriction Enzymes/metabolism Deoxyribonucleases, Type I Site-Specific Genetic Engineering/methods Genetic Vectors Histones/genetics Mutation Selection, Genetic

Chemicals

Histones DNA Restriction Enzymes endodeoxyribonuclease EcoK Deoxyribonucleases, Type I Site-Specific

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Waye M M

Verhoeyen M E

Jones P T

Winter G

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26 references, click to expand

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/13.23.8561

Published

1985-12-09

Pages

8561-71

Language

English

Region

England

NLM ID

0411011

PMCID

PMC322152

Subset

External Links

PubMed source DOI: 10.1093/nar/13.23.8561 PMC Full Text View journal details

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EcoK selection vectors for shotgun cloning into M13 and deletion mutagenesis.

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