Author: Lee, Na-Ra; Kwon, Hyun-Mi; Park, Kkothanahreum; Oh, Sangtaek; Jeong, Yong-Joo; Kim, Dong-Eun
Title: Cooperative translocation enhances the unwinding of duplex DNA by SARS coronavirus helicase nsP13 Document date: 2010_7_29
ID: 1k99yv4i_6
Snippet: SCV helicase nsP13 was overexpressed in E. coli and purified as described earlier (18) . DNA substrates were purchased from Integrated DNA Technologies (Coralville, IA, USA), and purified by a denaturing (8 M urea) polyacrylamide gel electrophoresis (PAGE). The DNA concentration was determined by 260 nm absorbance and its extinction coefficient. To produce duplex or triplex DNA substrates, the 32 P-labeled DNA strand was mixed with a 2-fold exces.....
Document: SCV helicase nsP13 was overexpressed in E. coli and purified as described earlier (18) . DNA substrates were purchased from Integrated DNA Technologies (Coralville, IA, USA), and purified by a denaturing (8 M urea) polyacrylamide gel electrophoresis (PAGE). The DNA concentration was determined by 260 nm absorbance and its extinction coefficient. To produce duplex or triplex DNA substrates, the 32 P-labeled DNA strand was mixed with a 2-fold excess of complementary DNA and then annealed by heating to 95 C followed by slowly cooling. The DNA substrates used in this study are listed in Table 1 .
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