Author： Xu Zhaohui
Publisher： Springer Publishing Company
ISSN： 1431-0651
Source： Extremophiles, Vol.15, Iss.6, 2011-11, pp. : 665-672
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Abstract
A putative Type II restriction-modification system of Thermotoga neapolitana</i>, Tne</i>DI, was cloned into Escherichia coli</i> XL1-Blue MRF′ and characterized. Gene CTN_0339 specifies the endonuclease R.Tne</i>DI, while CTN_0340 encodes the cognate DNA methyltransferase M.Tne</i>DI. Both enzymes were purified simply by heating the cell lysates of E. coli</i> 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.Tne</i>DI cleaved at the center of the recognition sequence (CG↓CG) and generated blunt-end cuts. Overexpression of R.Tne</i>DI in BL21(DE3) was confirmed by both SDS-PAGE and Western blotting.
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