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PEDS Advance Access published online on January 13, 2006
Protein Engineering Design and Selection, doi:10.1093/protein/gzj008
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© The Author 2006. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org
Received October 5, 2005
Revised December 7, 2005
Accepted December 8, 2005

Short Communication

Frame shuffling: a novel method for in vitro protein evolution

Kenji Kashiwagi 1, Yasuhiro Isogai 2, Kei-ichi Nishiguchi 3, and Kiyotaka Shiba 1 *

1 Department of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, 3-10-6, Ariake, Koto-ku, Tokyo 135-8550, Japan; CREST, Japanese Science and Technology Corporation Agency, Institute of Physical and Chemical Research (RIKEN), RIKEN Harima Institute, Mikazuki-cho, Sayo, Hyogo 679-5143, Japan
2 Biophysical Chemistry Laboratory, Institute of Physical and Chemical Research (RIKEN), RIKEN Harima Institute, Mikazuki-cho, Sayo, Hyogo 679-5143, Japan
3 Department of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, 3-10-6, Ariake, Koto-ku, Tokyo 135-8550, Japan

* To whom correspondence should be addressed.
Kiyotaka Shiba, E-mail: kshiba{at}jfcr.or.jp


  Abstract

We describe ‘frame shuffling’, a novel method for preparing artificial protein libraries. With this method, a Y-family DNA polymerase known to introduce frame shift mutations at high rates is utilized to scramble the reading frames of a parental gene. The resultant progeny produce mutant proteins having segmental sequence changes. Such frame-shuffled mutant proteins exhibit physicochemical properties that differ from those of proteins obtained using conventional mutagenesis.

Keywords: artificial protein; frame shift mutation; in vitro evolution; microgene; Y-family DNA polymerase.
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