caa a22 4500 445818387 CHVBK 20180317145236.0 cr unu---uuuuu 170323e20111101xx s 000 0 eng 10.1007/s00792-011-0397-9 doi (NATIONALLICENCE)springer-10.1007/s00792-011-0397-9 Cloning and characterization of the Tne DI restriction-modification system of Thermotoga neapolitana [Elektronische Daten] [Zhaohui Xu, Dongmei Han, Jingjing Cao, Uksha Saini] A putative Type II restriction-modification system of Thermotoga neapolitana, TneDI, was cloned into Escherichia coli XL1-Blue MRF′ and characterized. Gene CTN_0339 specifies the endonuclease R.TneDI, while CTN_0340 encodes the cognate DNA methyltransferase M.TneDI. Both enzymes were purified simply by heating the cell lysates of E. coli followed by centrifugation. The enzymes were active over a broad range of temperatures, from 42°C to at least 77°C, with the highest activities observed at 77°C. R.TneDI cleaved at the center of the recognition sequence (CG↓CG) and generated blunt-end cuts. Overexpression of R.TneDI in BL21(DE3) was confirmed by both SDS-PAGE and Western blotting. Springer, 2011 Thermotoga nationallicence Tne DI nationallicence Restriction-modification systems nationallicence Type II endonucleases nationallicence Xu Zhaohui Department of Biological Sciences, Bowling Green State University, 43403-0208, Bowling Green, OH, USA aut Han Dongmei Department of Biological Sciences, Bowling Green State University, 43403-0208, Bowling Green, OH, USA aut Cao Jingjing Department of Biological Sciences, Bowling Green State University, 43403-0208, Bowling Green, OH, USA aut Saini Uksha Department of Biological Sciences, Bowling Green State University, 43403-0208, Bowling Green, OH, USA aut Extremophiles Springer Japan 15/6(2011-11-01), 665-672 1431-0651 15:6<665 2011 15 792 https://doi.org/10.1007/s00792-011-0397-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00792-011-0397-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Xu Zhaohui Department of Biological Sciences, Bowling Green State University, 43403-0208, Bowling Green, OH, USA aut NATIONALLICENCE 700 1- Han Dongmei Department of Biological Sciences, Bowling Green State University, 43403-0208, Bowling Green, OH, USA aut NATIONALLICENCE 700 1- Cao Jingjing Department of Biological Sciences, Bowling Green State University, 43403-0208, Bowling Green, OH, USA aut NATIONALLICENCE 700 1- Saini Uksha Department of Biological Sciences, Bowling Green State University, 43403-0208, Bowling Green, OH, USA aut NATIONALLICENCE 773 0- Extremophiles Springer Japan 15/6(2011-11-01), 665-672 1431-0651 15:6<665 2011 15 792 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer