安徽农业科学2011，Vol.39Issue(17)：10153-10155,3.
Taq DNA聚合酶的热纯化制备
Preparation of Taq DNA Polymerase by Thermal Purification
摘要
Abstract
[ Objective ] The paper was to improve the preparation efficacy of Taq DNA polymerase. [ Method ] Ni column was used to purify Taq DNA polymerase carrying with 6xHis tag, and recombined vector. Using the thermal -resistant characteristics of TaqDNA polymerase, the crude extract was treated at 75 ℃ for 1 h, and the activity of prepared enzyme solution was verified by PCR test. [ Result] The recombinant pET-32A-Taq could highly express in BL21 (DE3) host bacteria and remove hybrid protein by thermal denaturation. The enzyme preparation with the activity further higher than purchased TaqDNA polymerase was obtained. [ Conclusion ] Taq DNA polymerase prepared by thermal purification method is simple with low cost, and can meet the needs of a large number of conventional PCR amplification.关键词
Taq DNA聚合酶/热纯化/pET-32A/BL21(DE3)Key words
Taq DNA polymerase/Thermal purification/pET-32A/BL21 (DE3)分类
生物科学引用本文复制引用
丁燕华,刘树涛,齐庆远..Taq DNA聚合酶的热纯化制备[J].安徽农业科学,2011,39(17):10153-10155,3.基金项目
国家自然科学基金委项目(30872254). （30872254）
