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PEDS Advance Access originally published online on January 13, 2006
Protein Engineering Design and Selection 2006 19(3):135-140; 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

SHORT COMMUNICATION

Frame shuffling: a novel method for in vitro protein evolution

Kenji Kashiwagi1,2, Yasuhiro Isogai3, Kei-Ichi Nishiguchi1 and Kiyotaka Shiba1,2,4

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

4 To whom correspondence should be addressed.E-mail: kshiba{at}jfcr.or.jp

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

Received October 5, 2005; revised December 7, 2005; accepted December 8, 2005.

Edited by Taiji Imoto


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