Plasmid and bacteriophage vectors for excision of intact inserts

Abstract

Plasmid (pPolyIII) and bacteriophage lambda (EMBL301) vectors are described in which sites for the rare-cutting enzymes SfiI and NotI (8-bp, recognition sequences) flank the polylinker cloning region. Intact DNA inserts for introduction into cultured cells or into the early embryo are readily excised from the vectors. General-purpose miniplasmid cloning vectors pPolyI and pPolyII are also described, and the utility of the bacteriophage lambda vector is demonstrated in the construction of a bovine genomic library.