定向进化
DNA洗牌
寡核苷酸
基因
重组
枯草芽孢杆菌
突变体
遗传学
FLP-FRT重组
生物
脂肪酶
计算生物学
化学
遗传重组
酶
生物化学
细菌
作者
Dongxing Zha,Andreas Eipper,Manfred T. Reetz
出处
期刊:ChemBioChem
[Wiley]
日期:2003-01-03
卷期号:4 (1): 34-39
被引量:93
标识
DOI:10.1002/cbic.200390011
摘要
A new and practical method for gene recombination with formation of libraries of mutant genes is presented. The method is based on the assembly of appropriately prepared oligonucleotides whose design is guided by sequence information. High recombination frequency with formation of full-length products is achieved by controlled overlapping of the designed oligomers. This process (ADO) minimizes self-hybridization of parental genes, which constitutes a significant advantage over conventional family shuffling as used in the directed evolution of functional enzymes. ADO was applied to the recombination of two lipase family genes from Bacillus subtilis (LipA and LipB). In a library of 3000 lipase variants created by this method, several were found that display increased enantioselectivity in a model reaction involving the hydrolysis of a meso-diacetate.
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