Arakawa Hiroshi, Kudo Hiroaki, Batrak Vera, Caldwell Randolph B, Rieger Michael A, Ellwart Joachim W, Buerstedde Jean-Marie
Institute for Molecular Radiobiology, Institute of Stem Cell Research, GSF-National Research Center for Environment and Health, Ingolstaedter Landstrasse 1, D-85764 Neuherberg-Munich, Germany.
Nucleic Acids Res. 2008 Jan;36(1):e1. doi: 10.1093/nar/gkm616. Epub 2007 Dec 11.
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.
群体内的全基因组突变和选择是自然进化的基础。当免疫球蛋白基因发生高突变，且只有那些表达抗原结合特异性提高的抗体的B细胞得以扩增时，抗体亲和力成熟过程中会发生类似的过程。如果转基因特异性高突变能够与携带有益突变的细胞选择相结合，那么蛋白质进化或许可以在细胞培养中模拟。在此，我们描述了通过高突变和迭代荧光激活细胞分选对B细胞系DT40中的绿色荧光蛋白（GFP）转基因进行优化。DT40中的人工进化具有独特优势，并且如果对理想突变的选择可行，那么它可以很容易地应用于其他转基因。
