易错PCR在酶的非理性设计改造中的应用
摘要
易错PCR是蛋白质分子改造非理性设计常用的技术,已被广泛应用于各类酶分子的性质改良。本文对易错PCR的基本原理及应用优势作了简要阐述,并总结了影响易错PCR效果的关键因素及近年来运用易错PCR技术改善酶的性质方面的研究进展。
引文
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[3]张秀艳.β-葡聚糖酶的定向进化及热稳定性研究[D].杭州:浙江大学, 2006.
[4]Cadwell R C, Joyce G F. Mutagenic PCR[J].Genome Research, 1992.
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[6]李文杰.枯草芽孢杆菌尿酸氧化酶的定向进化[D].中国科学技术大学,2017.
[7]Tang Z, Jin W, Rong S, et al. Improved thermostability and enzyme activity of a recombinant phyA mutant phytase from Aspergillus niger N25 by directed evolution and site-directed mutagenesis[J]. Enzyme Microb Technol, 2018, 108:74-81.
[8], Dagmara. Modification of fatty acid selectivity ofCandida antarcticalipase A by error-prone PCR[J]. Biotechnology Letters, 2017, 39(5):767-773.
[9]Fong S, Machajewski T D, Mak C C, et al. Directed evolution of D-2-keto-3-deoxy-6-phosphogluconate aldolase to new variants for the efficient synthesis of D-and L-sugars[J].Chemistry&Biology, 2000, 7(11):873-883.
[2]袁勤生.酶与酶工程(第二版)[M].上海:华东理工大学出版社,2012.
[3]张秀艳.β-葡聚糖酶的定向进化及热稳定性研究[D].杭州:浙江大学, 2006.
[4]Cadwell R C, Joyce G F. Mutagenic PCR[J].Genome Research, 1992.
[5]乔沛.通过定向进化和半理性设计提高顺式环氧琥珀酸水解酶的热稳定性的研究[D].杭州:浙江大学, 2016.
[6]李文杰.枯草芽孢杆菌尿酸氧化酶的定向进化[D].中国科学技术大学,2017.
[7]Tang Z, Jin W, Rong S, et al. Improved thermostability and enzyme activity of a recombinant phyA mutant phytase from Aspergillus niger N25 by directed evolution and site-directed mutagenesis[J]. Enzyme Microb Technol, 2018, 108:74-81.
[8], Dagmara. Modification of fatty acid selectivity ofCandida antarcticalipase A by error-prone PCR[J]. Biotechnology Letters, 2017, 39(5):767-773.
[9]Fong S, Machajewski T D, Mak C C, et al. Directed evolution of D-2-keto-3-deoxy-6-phosphogluconate aldolase to new variants for the efficient synthesis of D-and L-sugars[J].Chemistry&Biology, 2000, 7(11):873-883.