文献类型：期刊文献

英文题名：Mutant library construction in directed molecular evolution

作者：Wang, Tian-Wen; Zhu, Hu; Ma, Xing-Yuan; Zhang, Ting; Ma, Yu-Shu; Wei, Dong-Zhi

机构：[1]E China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2006

卷号：34

期号：1

起止页码：55

外文期刊名：MOLECULAR BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000240214900007)】；

语种：英文

外文关键词：directed molecular evolution; error-prone PCR; oligonucleotide-based mutagenesis; DNA shuffling; mutator strain; somatic hypermutation

摘要：Directed molecular evolution imitates the natural selection process in the laboratory to find mutant proteins with improved properties in the expected aspects by exploring the encoding sequence space. The success of directed molecular evolution experiment depends on the quality of artificially prepared mutant libraries and the availability of convenient high-throughput screening methods. Well-prepared libraries promise the possibility of obtaining desired mutants by screening a library containing a relatively small number of mutants. This article summarizes and reviews the currently available methodologies widely used in directed evolution practices in the hope of providing a general reference for library construction. These methods include error-prone polymerase chain reaction (epPCR), oligonucleotide-based mutagenesis, and genetic recombination exemplified by DNA shuffling and its derivatives. Another designed method is also discussed, in which B-lymphocytes are fooled to mutate nonantibody foreign proteins through somatic hypermutation (SHM).