摘要
目的制备重组Taq DNA聚合酶,为PCR提供试剂。方法用Taq DNA聚合酶基因的pTaq表达质粒转化E.coli菌株,异丙基硫代-β-D-半乳糖苷(IPTG)诱导12 h表达Taq DNA聚合酶,溶菌酶、NP40裂解细菌,硫酸铵沉淀、4℃透析,SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和PCR扩增分析其纯度和活性。结果分离纯化制备的Taq酶,纯度、活性都可与同类产品相比,能有效扩增DNA片段。结论该方法制备可用于PCR的Taq酶具有快速简便的优点。
Objective To prepare recombinant Taq DNA polymerase for PCR. Methods Taq DNA polymerase was expressed in recombinant E. coli strain under isopropy-β-D-thiogalactoside （IPTG） induction for 12 h, disrupted with lysozyme and NP 40, followed dialyzed at 4 ℃. Purification and activities of polymerase were analyzed under sodium dodecyl sulfate polyacrylamide gel electropheresis （SDS-PAGE） and polymerase chain reaction（PCR） methods. Results The prepared polymerase purification and activities could compare with the same product can be used to amplify the DNA fragment. Conclusion The prepared methods of recombinant Taq DNA polymerase in our laboratory are simple and rapidly.
出处
《新乡医学院学报》
CAS
2007年第6期551-553,共3页
Journal of Xinxiang Medical University
基金
河南省科技厅自然科学基金(编号:511042300)
河南省科技攻关资助项目(编号:624410041)
关键词
TAQ
DNA聚合酶
基因工程
分离纯化
Taq DNA polymerase
gene engineering
separation and purification
