λ ZAP: a bacteriophage λ expression vector with in vivo excision properties

摘要: A lambda insertion type cDNA cloning vector, Lambda ZAP, has been constructed. In E. coli a phagemid, pBluescript SK(-), contained within the can be excised by f1 or M13 helper phage. The excision process eliminates need to subclone DNA inserts from phage into plasmid restriction digestion and ligation. This is possible because ZAP incorporates signals for both initiation termination of synthesis bacteriophage origin replication (1). Six 21 sites in SK polylinker, NH2-portion lacZ gene, are unique ZAP. Coding sequences inserted these sites, appropriate reading frame, expressed promoter as fusion proteins. features this vector significantly increase rate at which clones isolated analyzed. was tested preparation chicken liver library isolation actin screening with oligonucleotide probes. Putative were identified sequencing. ability serve construction expression libraries determined detecting proteins containing glucocerbrosidase cDNA's using rabbit IgG anti-glucocerbrosidase antibodies.