A modified pBR322 vector with improved properties for the cloning, recovery, and sequencing of blunt-ended DNA fragments.
The construction of a plasmid vector which facilitates the cloning and recovery of blunt-ended DNA fragments is described. This plasmid, called pHP34, differs from pBR322 by a 10-bp insertion which introduces a unique SmaI site immediately flanked by two EcoRI sites. Blunt-ended DNA fragments cloned in the SmaI site can be recovered by digestion with EcoRI. Small cloned fragments can be chemically sequenced using a strategy which does not require their purification. The use of a plasmid related to pHP34 for in vitro mutagenesis by the insertion of a DNA linker fragment conferring an antibiotic resistance is also discussed.
D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D009154:Mutation; D010641:Phenotype; D010957:Plasmids; D011995:Recombination, Genetic
D004269:DNA, Bacterial