体外定向分子进化的新策略及新应用

体外定向分子进化是发现和改造生物活性分子的重要方法,提供了一种高效的获得多样性的方法。DNA改组(DNA shuffling)是重要的体外分子进化技术,结合高通量筛选能够改造许多重要的医药、工业、环境保护等方面的商业酶。近年来,许多体外分子进化的新策略和新方法层出不穷,得到了良好的发展和应用,其中有代表性的11种是DNA家族改组(DNA family shuffling),部分基因片段改组、单链DNA家族改组(SSDNAs)、简并引物基因改组(DOGS)、基因组改组(Genome Shuffling)、瞬时模板的随机嵌合(RACHITT)、单向引物的随机重组(MURA)、自我复制(CSR)改组、易错环行扩增(error-prone RCA)、基于遗传密码随机切除(COBARDE)、核酸内切酶V(endonuclease V)替代核酸内切酶DNaseI等。

New Strategies and New Application of Molecular in vitro Evolution

Molecular in vitro evolution is a powerful engine for the creation of new phenotypes. Many useful enzymes and peptides have been isolated following the artificial evolution. An exciting development was the shuffling to create diversity for directed evolution. DNA shuffling is an important technology for in vitro directed molecular evolution. DNA shuffling and high throughput screening were a powerful tool for optimization of many commercially important enzymes for which selections do not exist. The uses of enzymes were applied to medical, industrial and environmental purposes. A number of new different mutagenesis strategies exist, such as DNA family shuffling, shuffling of critical segments of gene, shuffling of single-stranded DNA, Degenerate oligonucleotide gene shuffling, Genome Shuffling, random chimeragenesis on transient templates, mutagenic and unidirectional reassembly, compartmentalized self-replication, error-prone rolling circle amplification, codon-based random deletion,endonuclease V etc.