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GENERAL SELECTION FOR SPECIFIC DNA-BINDING ACTIVITIES
Nicholas Benson 1, Paul Sugiono 1, Steven Bass 1, Lynn V. Mendelman 1, and Philip Youderian 1
1 Department of Biological Sciences, University of Southern
California, Los Angeles, California 90089-1481
We present a general strategy for the selection of bacterial
clones that express DNA-binding activities corresponding to particular DNA
recognition sites. The selection uses a "challenge phage" vector, P22
Kn9 arc-amH1605, into which is substituted a synthetic DNA-binding site
for a site that controls transcription of the P22 antirepressor (ant)
gene. Constitutive synthesis of antirepressor channels a challenge phage into
lytic development and efficiently kills an infected host, unless the substituted
site is bound by a specific protein; in this case, the challenge phage prefers
lysogenic development, and the host survives and acquires an antibiotic-resistance
phenotype. Infections with challenge phages carrying the E. coli Lac
operator, phage 
OL1 operator, or synthetic, "idealized"
E. coli Trp and Tn10 Tet operators select clones that express
each of the corresponding binding activities. The use of challenge phage vectors
may be extended to select clones that express eukaryotic DNA-binding activities.
Accepted on May 7, 1986
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