id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-275016-ij5yaqkx	Someya, Yuichi	Characterization of the norovirus 3C-like protease	2005-03-08		.txt	text/plain	3244	187	60	The recombinant 3C-like protease of Chiba virus, a Norovirus, expressed in Escherichia coli cells was purified and characterized as to effects of pH, temperature, salt contents, and SH reagents on its proteolytic activity. The DNA fragment encoding all residues (Ala1 to Glu181) of the Chiba virus 3C-like protease was amplified by PCR, in that NdeI and Aor51HI restriction sites were introduced at the 5 and 3 ends, respectively. In order to observe proteolysis at the cleavage site between the 3C and 3D, we at first used the His-3CD-C139A mutant protein expressed from pT7His3CD-C139A as a substrate, which was the N-terminal His-tagged 3CD fragment containing the Ala mutation of active-site Cys139 of the 3C-like protease (Fig. 1B) . They used bacterially expressed 3C-like protease with its C-terminus His-tagged as an enzyme and the entire ORF1 protein or 3CD fragment containing the active-site Cys mutation as a substrate which was expressed in the in vitro transcription/translation reaction.	./cache/cord-275016-ij5yaqkx.txt	./txt/cord-275016-ij5yaqkx.txt