Abstract
λgt11 is a 43.7-kb linear double-stranded λ bacteriophage cloning vector designed for cloning small Eco RI fragments, particularly cDNA fragments with Eco RI linkers. The unique E co Rl site is used for inserting foreign DNA, which is located near the COOH terminus of the lac Z gene. Foreign DNA sequences in this cloning vehicle are expressed as β-galactosidase fusion proteins. Recombinant libraries generated from λgt11 are screened with either antibody probes or nucleic acid probes, because the antibody detects the fusion protein expressed by the recombinant λgt11 bacteriophage. Insertion of foreign DNA into the Eco RI site renders the bacteriophage gal – , whereas nonrecombinant λgt11 is gal + . This chapter discusses the two useful genetic properties of the cloning vehicle: (1) it contains a temperature-sensitive repressor of lytic growth that is inactive at 42°C and active at 32°C, allowing a selection against lysogenic growth, and (2) it has an amber mutation, S 100, that renders the phage host lysis defective in all E. Coli hosts that do not contain a strong amber suppression mutation.
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