A bacterial two-hybrid selection system for studying protein-DNA and protein-protein interactions.
We have developed a bacterial "two-hybrid" system that readily allows selection from libraries larger than 10(8) in size. Our bacterial system may be used to study either protein-DNA or protein-protein interactions, and it offers a number of potentially significant advantages over existing yeast-based one-hybrid and two-hybrid methods. We tested our system by selecting zinc finger variants (from a large randomized library) that bind tightly and specifically to desired DNA target sites. Our method allows sequence-specific zinc fingers to be isolated in a single selection step, and thus it should be more rapid than phage display strategies that typically require multiple enrichment/amplification cycles. Given the large library sizes our bacterial-based selection system can handle, this method should provide a powerful tool for identifying and optimizing protein-DNA and protein-protein interactions.
Joung
J K
JK
Howard Hughes Medical Institute and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Ramm
E I
EI
Pabo
C O
CO
D001419:Bacteria; D001681:Biological Assay; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D015723:Gene Library; D019151:Peptide Library; D011485:Protein Binding; D011506:Proteins / Q000032:analysis* / Q000235:genetics*
D019151:Peptide Library; D011506:Proteins; D004247:DNA