External Surface Display of Proteins Linked to DNA-Binding Domains

✍ Scribed by Duncan P. McGregor; Simon P. Robins

• Publisher: Elsevier Science
• Year: 2001
• Tongue: English
• Weight: 176 KB
• Volume: 294
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2001.5147

✦ Synopsis

A novel system (DBDX) was developed which allows the external surface display on filamentous bacteriophage of proteins fused to either the N- or the C-terminus of a DNA-binding protein. In conjunction with helper phage infection, expression of proteins fused to the estrogen receptor DNA-binding domain (DBD) in a phagemid vector containing the DNA sequence recognized by the DBD resulted in the production of phage particles which display the fusion protein through the phage pVIII coat on the external surface of the particle. The viability of the technique was established with several model systems: particles displaying the C-terminal domain of N-cadherin or the biotinylation domain of propionyl coenzyme A carboxylase fused to the C-terminus of the DBD were found to be bound specifically by antibody or streptavidin, respectively. Human kappa constant region cDNA was selected from a N-terminal DBD fusion lymphocyte cDNA library after two rounds of selection with anti-kappa antibody. This display system may complement currently available bacterial selection techniques.