pEZZ 18 Protein A Gene Fusion Vector
pEZZ 18 Protein A Gene Fusion Vector
pEZZ 18 Protein A Gene Fusion Vector
pEZZ 18 Protein A Gene Fusion Vector
概述
- For rapid expression of secreted fusion proteins and their one-step purification using IgG Sepharose 6 Fast Flow
The phagemid pEZZ 18 contains the protein A signal sequence and two synthetic “Z” domains based on the “B” IgG binding domain of protein A. Proteins are expressed as fusions with the “ZZ” peptide and secreted into the aqueous culture medium under the direction of the protein A signal sequence. They are easily purified using IgG Sepharose 6 Fast Flow to which the “ZZ” domain binds tightly. Because of its folding properties, the 14 kDa “ZZ” peptide has little effect on folding of the fusion partner into a native conformation.
Expression
Expression is controlled by the lacUV5 and protein A promoters and is not inducible. Elements of the protein A gene provide the ATG and ribosome-binding sites. Stop codons must be provided by the insert
Sequencing
M13 Universal Sequencing Primer can be used for both double-stranded and single-stranded sequencing. A protocol for production of single-stranded DNA is provided with the vector
Cloning
Inserts containing a stop codon will yield white colonies when grown on media containing X-gal
Host(s)
E. coli strains carrying a lac deletion but capable of a-complementation of lacZ’.