A novel cloning vector for the direct selection of recombinant DNA in E. coli.
A new plasmid cloning vector (pHE3) is described carrying dominant p-fluorophenylalanine-sensitivity (pheS) and chloramphenicol-resistance markers. This vector is used in combination with a p-fluorophenylalanine-resistant Escherichia coli recipient (strain RR28). Foreign DNA can be cloned into pHE3 leading to insertional inactivation of pheS. Transformation of RR28, and plating on minimal medium with chloramphenicol plus p-fluorophenylalanine (pfp) directly selects for colonies containing recombinant DNA.
D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D005799:Genes, Dominant; D010957:Plasmids; D014170:Transformation, Genetic; D010135:p-Fluorophenylalanine / Q000494:pharmacology
D004274:DNA, Recombinant; D010135:p-Fluorophenylalanine; D004262:DNA Restriction Enzymes