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Nucleic Acids Research, 1985, Vol. 13, No. 23 8561-8571
© 1985

Articles

EcoK selection vectors for shotgun cloning into M13 and deletion mutagenesls

Mary M.Y. Waye, Martine E. Verhoeyen, Peter T. Jones and Greg Winter

MRC Laboratory of Molecular Biology Hills Road, Cambridge CB2 2QH, UK

Received September 3, 1985. Revised November 5, 1985. Accepted November 5, 1985.

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.


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