title: EcoK selection vectors for shotgun cloning into M13 and deletion mutagenesis.
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: D001483:Base Sequence; D002872:Chromosome Deletion; D003001:Cloning, Molecular / Q000379:methods*; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D015253:Deoxyribonucleases, Type I Site-Specific; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D006657:Histones / Q000235:genetics; D009154:Mutation; D012641:Selection, Genetic
chemical_list: D006657:Histones; D004262:DNA Restriction Enzymes; C029602:endodeoxyribonuclease EcoK; D015253:Deoxyribonucleases, Type I Site-Specific