期刊
JOURNAL OF IMMUNOLOGICAL METHODS
卷 251, 期 1-2, 页码 187-193出版社
ELSEVIER
DOI: 10.1016/S0022-1759(01)00300-3
关键词
mutator cells; recombinant DNA; antibody; phage display; mutagenesis
This protocol provides a simple in vivo strategy for introducing random mutations to a target DNA sequences using E. coli mutator cells. The method has been used in our laboratory for affinity maturation of proteins encoded by target DNA sequences. Selection conditions can be modified for antibody fragments with increased production levels. Growth conditions in E. coli mutator cells can be adjusted to introduce a single random point mutation per kilobase of DNA, approximately equivalent to one codon change per scFv fragment. (C) 2001 Elsevier Science B.V. All rights reserved.
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