Wang J.,China Pharmaceutical University |
Wang J.,East China Research Institute for Medicine and Biotechnics |
Zou B.,China Pharmaceutical University |
Zou B.,East China Research Institute for Medicine and Biotechnics |
And 7 more authors.
Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology | Year: 2011
We expressed recombinant single-stranded DNA-binding protein (r-SSBP) from Escherichia coli with the molecular weight of 24-kDa by using genetic engineering strategy, and demonstrated the single-stranded DNA (ssDNA)-binding activity of r-SSBP by electrophoretic mobility shift assay (EMSA). To further characterize r-SSBP, we studied the effects of r-SSBP on melting temperature (Tm) of DNA. The results showed that r-SSBP could bind to ssDNA, and lower the Tm of DNA, especially for single-base mismatched DNA. Therefore, r-SSBP significantly increased the Tm difference between single-base mismatched DNA and perfect matched DNA. These results are very beneficial for single-nucleotide polymorphism detection. Moreover, we applied r-SSBP in high sensitive pyrosequencing system developed by our group. The results suggest that the r-SSBP decreased non-specific signals, corrected the proportion of signal peak height and improved the performance of pyrosequencing. © 2011 CJB, All rights reserved. Source