ATPase activity measurement of DNA replicative helicase from Bacillus stearothermophilus by malachite green method |
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| Authors: | Mu Yang Ganggang Wang |
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| Affiliation: | 1. Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, China;2. Key Laboratory of Environmental Microbiology of Sichuan Province, Chengdu, 610041, China;3. University of Chinese Academy of Sciences, Beijing, 100049, China |
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| Abstract: | The DnaB helicase from Bacillus stearothermophilus (DnaBBst) was a model protein for studying the bacterial DNA replication. In this work, a non-radioactive method for measuring ATPase activity of DnaBBst helicase was described. The working parameters and conditions were optimized. Furthermore, this method was applied to investigate effects of DnaG primase, ssDNA and helicase loader protein (DnaI) on ATPase activity of DnaBBst. Our results showed this method was sensitive and efficient. Moreover, it is suitable for the investigation of functional interaction between DnaB and related factors. |
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| Keywords: | ATPase activity DnaB Malachite green Optimization |
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