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I-PpoI and I-CreI Homing Site Sequence Degeneracy Determined by Random Mutagenesis and Sequential
 

Summary: I-PpoI and I-CreI Homing Site Sequence Degeneracy
Determined by Random Mutagenesis and Sequential
in Vitro Enrichment
Gretchen M. Argast1
, Kathryn M. Stephens1
, Mary J. Emond2
and Raymond J. Monnat Jr1
*
1
Departments of Pathology and
2
Biostatistics, University of
Washington, Seattle, WA 98195
USA
Plasmid libraries containing partially randomized cleavage sites for the
eukaryotic homing endonucleases I-PpoI and I-CreI were constructed, and
sites that could be cleaved by I-PpoI or I-CreI were selectively recovered
by successive cycles of cleavage and gel separation followed by religation
and growth in Escherichia coli. Twenty-one different I-PpoI-sensitive hom-
ing sites, including the native homing site, were isolated. These sites

  

Source: Aydin, Zafer - Department of Genome Sciences, University of Washington at Seattle

 

Collections: Biology and Medicine