CHARACTERIZATION OF RECOMBINANT SINGLE-STRAND BINDING PROTEIN FROM ESCHERICHIA COLI AND ITS APPLICATION IN PYROSEQUENCING
We expressed the recombinant single-strand binding protein (r-SSBP) from Escherichia coli with the molecular weight of 24-kDa by using the genetic engineering strategy. To characterize the r-SSBP, we studied the effects on melting temperature (Tm) of DNA. The results showed that the r-SSBP can bind to ssDNA, and lower the Tm of DNA, especially for single-base mismatched DNA. Therefore, the r-SSBP can significantly increase the Tm difference between single-base mismatched DNA and perfect matched DNA. These results are very beneficial for singlenucleotide polymorphism detection. Moreover, we applied the r-SSBP in high sensitive pyrosequencing system developed by our group. The results suggested that the r-SSBP can decrease unspecific signals and improve the performance of pyrosequencing. This research laid the foundation for the development of high sensitive pyrosequencing regents.
Escherichia coli single-strand binding protein melting temperature pyrosequencing
Wang Jianping Zou Bingjie Chen Zhiyao Ma Yinjiao Xu Shu Zhou Guohua
School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China Huadon Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China
国际会议
重庆
英文
256-259
2011-10-28（万方平台首次上网日期，不代表论文的发表时间）