ZAP Express System - Details & Specifications
The ZAP Express vector (see Figure 1) allows both eukaryotic and prokaryotic expression, while also increasing both cloning capacity and the number of unique lambda cloning sites. The ZAP Express vector has 12 unique cloning sites which will accommodate DNA inserts from 0 to 12 kb in length. The 12 unique cloning sites are Apa I, BamH I, EcoR I, Hind III, Kpn I, Not I, Sac I, Sal I, Sma I, Spe I, Xba I, and Xho I. Inserts cloned into the ZAP Express vector can be excised out of the phage in the form of the kanamycin-resistant pBK-CMV phagemid vector by the same excision mechanism found in the Lambda ZAP vectors.
Clones in the ZAP Express vector can be screened with either DNA probes or antibody probes, and in vivo rapid excision of the pBK-CMV phagemid vector allows insert characterization in a plasmid system. The polylinker of pBK-CMV phagemid has 17 unique cloning sites flanked by T3 and T7 promoters and has 5 primer sites for DNA sequencing. The plasmid has the bacteriophage f1 origin of replication allowing rescue of single-stranded DNA, which can be used for DNA sequencing or site-directed mutagenesis. Unidirectional deletions can be made using exonuclease III and mung bean nuclease by taking advantage of the unique positioning of 5´ and 3´ restriction sites. Transcripts made from the T3 and T7 promoters generate riboprobes useful in Southern and Northern blotting, and the lacZ promoter may be used to drive expression of fusion proteins suitable for Western blot analysis or protein purification.
Eukaryotic expression of inserts is driven by the cytomegalovirus (CMV) immediate early (IE) promoter with the SV40 transcription terminator and polyadenylation signal.
Stable selection of clones in eukaryotic cells is made possible by the presence of the neomycin- and kanamycin-resistance gene, which is driven by the SV40 early promoter with TK transcription polyadenylation signals to render transfectants resistant to G418 (geneticin).
For Research Use Only. Not for use in diagnostic procedures