EcoK selection vectors for shotgun cloning into M13 and deletion mutagenesls
- 1 January 1985
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 13 (23) , 8561-8571
- https://doi.org/10.1093/nar/13.23.8561
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.Keywords
This publication has 26 references indexed in Scilit:
- Studies on transformation of Escherichia coli with plasmidsPublished by Elsevier ,2006
- Membrane-bound domain of HMG CoA reductase is required for sterol-enhanced degradation of the enzymeCell, 1985
- Generation of β-globin by sequence-specific proteolysis of a hybrid protein produced in Escherichia coliNature, 1984
- Preliminary crystallographic study of the complex between the Fab fragment of a monoclonal anti-lysozyme antibody and its antigenJournal of Molecular Biology, 1983
- In VitroDeletional Mutagenesis for Bacterial Production of the 20,000-Dalton Form of Human Pituitary Growth Hormone.DNA, 1983
- The use of synthetic oligodeoxyribonucleotides to produce specific deletions in the araBAD promoter of Escherichia coli B/rGene, 1982
- Structures and mechanisms of DNA restriction and modification enzymesQuarterly Reviews of Biophysics, 1979
- The nucleotide sequence recognized by the Escherichia coli K12 restriction and modification enzymesJournal of Molecular Biology, 1979
- Complete sequence of constant and 3′ noncoding regions of an immunoglobulin mRNA using the dideoxynucleotide method of RNA sequencingCell, 1978
- Specific binding of a synthetic oligodeoxyribonucleotide to yeast cytochrome c mRNANature, 1977