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Nucleic Acids Research Advance Access originally published online on December 11, 2007
Nucleic Acids Research 2008 36(1):e1; doi:10.1093/nar/gkm616
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Nucleic Acids Research, 2008, Vol. 36, No. 1 e1
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

Protein evolution by hypermutation and selection in the B cell line DT40

Hiroshi Arakawa1, Hiroaki Kudo1, Vera Batrak1, Randolph B. Caldwell1, Michael A. Rieger2, Joachim W. Ellwart3 and Jean-Marie Buerstedde1,*

1Institute for Molecular Radiobiology, 2Institute of Stem Cell Research, GSF-National Research Center for Environment and Health, Ingolstaedter Landstrasse 1, D-85764 Neuherberg-Munich and 3Institute of Molecular Immunology, GSF National Research Center for Environment and Health, Marchioninistrasse 25, 81377 Munich, Germany

*To whom correspondence should be addressed. Tel: +49 89 3187 2871; Fax: +49 89 3187 4093; Email: buersted{at}gsf.de

Received May 24, 2007. Revised July 3, 2007. Accepted July 29, 2007.

Genome-wide mutations and selection within a population are the basis of natural evolution. A similar process occurs during antibody affinity maturation when immunoglobulin genes are hypermutated and only those B cells which express antibodies of improved antigen-binding specificity are expanded. Protein evolution might be simulated in cell culture, if transgene-specific hypermutation can be combined with the selection of cells carrying beneficial mutations. Here, we describe the optimization of a GFP transgene in the B cell line DT40 by hypermutation and iterative fluorescence activated cell sorting. Artificial evolution in DT40 offers unique advantages and may be easily adapted to other transgenes, if the selection for desirable mutations is feasible.

Present address: Hiroaki Kudo, Graduate School of Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan


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