43] In vivo excision properties of bacteriophage λ ZAP expression vectors

Abstract

Publisher Summary The λ ZAP vector system, although originally designed for the efficient construction and characterization of complementary DNA (cDNA) clones has proved useful for a broad range of applications. With an insert capacity of over 10 kb, the phage has also been used successfully for the construction of plant and bacterial genomic libraries. The construction of genomic libraries with smaller insert size is frequently beneficial in cloning a larger percentage of the genome in instances where large DNA clones containing unusual structure or secondary modifications are unstable. The requirement for producing a larger library to compensate for smaller insert size in order to obtain genome representation is frequently offset by the speed at which λ ZAP clones can be characterized. The chapter provides information about the modified λ ZAP vector system for the expression of antibody variable regions cloned via polymerase chain reaction (PCR) amplification. The chapter also discusses the introduction of a modified λ ZAP vector into the chromosome of transgenic mice for the purpose of generating a λ shuttle vector system capable of measuring spontaneous and induced tissue specific mutation rates in whole animals. The in vivo excision features of the mutagenesis λ ZAP vector permit rapid recovery of the target gene from the h phage for sequence identification of selected mutations.