Abstract
A thermostable DNA polymerase I from a mesophilic Bacillus sphaericus strain C3-41 was characterized in this study. The polI was cloned, sequenced and over-expressed in Escherichia coli. The expressed 110 kDa fusion protein of PolI was stable at 70°C for 1 h. Compared with DNA polymerase I of E. coli (TaKaRa), the relative polymerase activity of this PolI was 3.33 ± 0.1 RFU μl−1 at 37°C using fluorescent quantitative analysis. It showed higher polymerase activity than E. coli PolI at higher temperature, with a relative activity of 3.75 ± 0.1 RFU μl−1 at 70°C. The polI sequence analysis and the protein structure prediction indicated that this protein had a high similarly to other PolI from thermophilic micro-organisms. This information is of importance for future study for evolution of the house-keeping gene polI in entomopathogenic bacterium B. sphaericus.
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Acknowledgments
We thank Dr. Simon Rayner for useful suggestion and critical reading of the manuscript, and Mr. Cai Quanxin for his technical assistance. The project was supported by grants (KSCX2-SW-301-10, KSCX2-SW-315) from the Chinese Academy of Sciences, a 973 project (2003CB114201) and a grant (30470037) from NFSC, China.
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Bei, H., Haizhou, L., Xiaomin, H. et al. Preliminary characterization of a thermostable DNA polymerase I from a mesophilic Bacillus sphaericus strain C3-41. Arch Microbiol 186, 203–209 (2006). https://doi.org/10.1007/s00203-006-0135-3
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DOI: https://doi.org/10.1007/s00203-006-0135-3