Vector displays

Fig. 5.3. Schematic representation of die display vector pASKIntlOO. fl, fl replication origin cat, chloramphenicol resistance marker tetR, tetracycline repressor encoding gene tetP/O, tetracycline promotor/operator region colEl, ColEl replication origin intimin, truncated eaeA gene of EHEC 0157 H7. Unique Ava I (Sma I, Xma I) and Bam HI restriction sites allow die in-frame fusion of genes encoding various passenger domains, as described in furdier detail in Wentzel et al. [7].

Phage Display Vectors for Cloning of Antibody Genes in Alphabetical Order co... 

Phage display vector Promoter Secretion Antibody format used C-domains ir vector Sites heavy chali Sites light chain Tags gill Expression of soluble Ab Reference... 

Tsurushita, N., Fu, H., and Warren, C. (1996) Phage display vectors for in vivo recombination of immunoglobulin heavy and light chain genes to make large combinatorial libraries. Gene 172, 59-63. 

Phage Display Vectors for the In Vitro Generation of Human... 

A large number of different phage display vectors have been constructed. With pretending to be complete, Table 1 lists a selection of phage display vectors. Some of them have not been used for the construction of a library up to now but have been included because they offer possible alternatives. For example, one of the systems allows the success of antibody gene cloning to be monitored by the expression of green fluorescent protein (52). 

Den, W., Sompuram, S. R., Sarantopoulos, S., and Sharon, J. (1999) A bidirectional phage display vector for the selection and mass transfer of polyclonal antibody libraries. J. Immunol. Meth. 222, 45-57. 

NOMENCLATURE FOR M13 TYPE PHAGE- AND PHAGEMID-DISPLAY VECTOR TYPES AS PROPOSED BY SMITH [209]... 

The various modalities in which proteins are presented on the display vector are illustrated in Fig. 3 with respect to the association of monomers, heterodimers, and particularly for antibodies, the Fab, scFv and diabody display (see Refs. 38 and 39 for reviews). The direct interaction rescue system is treated separately below. 

Two other bacteriophage systems have also been investigated as potential phage-display vectors, namely bacteriophage T4 [131], where a C-terminal extension on fibritin encoded by gene wac (whisker s antigen control) could be displayed, and P4 [132], where a deca-peptide was successfully inserted and presented near the N-terminus of the capsid decoration component, Psu. Their relative usefulness cannot yet be evaluated. 

Efimov YP, Nepluev IY, Mesyanzhinov VV, Bacteriophage T4 as a surface display vector, Virus Genes, 10 173-177, 1995. 

Markland W, Roberts BL, Saxena MJ, Guterman SK, Ladner RC, Design, construction and function of a multicopy display vector using fusion to the major coat protein of bacteriophage M13, Gene, 109 13-19, 1991. 

Display on lambda phages has been described with two major coat proteins the gpV tail protein [23, 24] and the gpD head protein [25, 26]. Both proteins accept C-terminal fusions, which offers alternative systems for displaying cDNA libraries. A high level of display is possible as demonstrated by the total modification of gpV [27] and gpD [25] proteins. The structure of gpD has been solved recently [28] this should help the design of optimal display vectors. 

In phagemid vectors, a stop codon is often introduced between the foreign protein and the phage coat protein. Hence, display is only possible when the infected E.coli strain contains an appropriate suppressor. Such a phage display vector behaves as a classical expression vector when a non-suppressor strain is transformed. This system eliminates the need for a recloning step for the production of the free enzyme after selection. Nevertheless, the risk of obtaining a very low level of surface expression is increased as suppression is always partial. 

---