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

被引：35

作者：

Arakawa, Hiroshi ^{[1
]}

Kudo, Hiroaki ^{[1
]}

Batrak, Vera ^{[1
]}

Caldwell, Randolph B. ^{[1
]}

Rieger, Michael A. ^{[2
]}

Ellwart, Joachim W. ^{[3
]}

Buerstedde, Jean-Marie ^{[1
]}

机构：

[1] GSF Natl Res Ctr Environm & Hlth, Inst Mol Radiobiol, D-85764 Neuherberg, Germany

[2] GSF Natl Res Ctr Environm & Hlth, Inst Stem Cell Res, D-85764 Neuherberg, Germany

[3] GSF Natl Res Ctr Environm & Hlth, Inst Mol Immunol, D-81377 Munich, Germany

来源：

NUCLEIC ACIDS RESEARCH | 2008年 / 36卷 / 01期

关键词：

D O I：

10.1093/nar/gkm616

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页数：11

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