Phage Lambda and Molecular Cloning

摘要: For more than 20 years, transducing derivatives of lambdoid phages have facilitated functional and structural analyses bacterial genes. Restriction enzymes that make staggered cuts within specific DNA sequences (targets) produce discrete fragments with short cohesive ends (Hedgpeth et al. 1972; Mertz Davis Bigger 1973), this discovery immediately indicated could be spliced into the severed arms a λ vector molecule. The cohered would then joined covalently by ligase (Gellert 1967), recombinant genomes, phages, recovered transfection Escherichia coli (Mandel Higa 1970). This goal was quickly realized (Murray Murray 1974; Thomas 1974), detailed type previously possible for only few E. genes became generally applicable. ability to insert from any source plasmid phage vectors, supplemented new techniques rapid sequence determination (Maxam Gilbert 1977; Sanger 1977), has already generated many exciting advances in diverse areas molecular biology. development chromosome as receptor assured availability well-characterized deletion mutants, means selecting loss restriction targets prior identification essential components transcriptional circuits replication system. precise knowledge attained 1972 showed genetic engineer which regions genome sacrificed and, also, special features be...